CN110300520A - Anti- C1s antibody and its application method - Google Patents

Anti- C1s antibody and its application method Download PDF

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CN110300520A
CN110300520A CN201780074920.XA CN201780074920A CN110300520A CN 110300520 A CN110300520 A CN 110300520A CN 201780074920 A CN201780074920 A CN 201780074920A CN 110300520 A CN110300520 A CN 110300520A
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CN110300520B (en
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S·帕尼克
G·帕瑞
N·E·斯坦格莉安奥
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United States Than Aoweiladiwei Ltd By Share Ltd
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    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
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    • C07K2317/52—Constant or Fc region; Isotype
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    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
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    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
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    • C07K2317/00—Immunoglobulins specific features
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    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

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Abstract

The antibody of disclosure offer specific binding complement pathway components C1s.The disclosure provides the nucleic acid of the nucleotide sequence comprising encoding the anti-C1s antibody；With the host cell comprising the nucleic acid.The disclosure provides the composition comprising the anti-C1s antibody.The disclosure provides the method for using the anti-C1s antibody.

Description

Anti- C1s antibody and its application method
The intersection of related application is quoted
The equity of the U.S. Provisional Application for the serial number 62/407,390 that this application requires on October 12nd, 2016 to submit, leads to It crosses to quote and is completely included in this article it.
The sequence table submitted to electronics is quoted
This application contains ordered list, has been submitted via EFS-Web with ASCII fromat, and accordingly by having quoted It is whole to include.The ASCII copy of creation on October 11st, 2017 is named as 4159_504PC01_SeqListing_ ST25.TXT and size are 72,260 bytes.
Background of invention
Complement system is a kind of well known effector mechanism of immune response, not only provides and is directed to pathogen and other nocuousness The protection of substance, and provide from injury recovery.Complement pathway includes typically existing more in the body with inactive form Kind albumen.Classic complement approach by complement first composition (referred to as C1 compound, by C1q, C1r, and C1s albumen form) work Change triggering.Once C1 combination immune complex or other activators, a kind of C1s ingredient (diisopropyl fluorophosphate (DFP) sensibility Serine protease) complement component C4 and C2 are cut to start the activation of classic complement approach.Classic complement approach seems permitted It plays a role in more diseases and illness.
Summary of the invention
The disclosure provides the anti-C1s antibody of humanization.The disclosure provides the nucleotide comprising the coding anti-C1s antibody of the humanization The nucleic acid of sequence；With the host cell comprising the nucleic acid.The disclosure provides the composition comprising the anti-C1s antibody of the humanization.This It is open that the method for using the anti-C1s antibody of the humanization is provided.
The some aspects of the disclosure are related to a kind of antibody, include heavy chain and light chain, wherein the heavy chain includes weight chain variable (VH) area and heavy chain constant region, the light chain include the area light chain variable (VL)；Wherein the area VL includes VL complementary determining region (CDR) 1, VL CDR2, and VL CDR3, and wherein the area VH includes VH CDR1, VH CDR2, and VH CDR3；The wherein VL CDR1 packet The NO:1 of ID containing SEQ；Wherein the VL CDR2 includes SEQ ID NO:2；Wherein the VL CDR3 includes SEQ ID NO:3；Wherein The VH CDR1 includes SEQ ID NO:4；Wherein the VH CDR2 includes SEQ ID NO:5；Wherein the VH CDR3 includes SEQ ID NO:6；Wherein the heavy chain constant region includes IgG4 constant region, wherein the amino of the heavy chain constant region corresponding to SEQ ID NO:28 Sour residue 308 is Leu, and the amino acid residue 314 of the heavy chain constant region corresponding to SEQ ID NO:28 is Ser；And wherein should Antibody specificity combines the C1s of activation.
The some aspects of the disclosure are related to a kind of antibody, include heavy chain and light chain, wherein the heavy chain includes the area VH and heavy chain Constant region, and the light chain include the area VL；Wherein the area VL includes VL CDR1, VL CDR2, and VL CDR3, and the wherein area VH Include VH CDR1, VH CDR2, and VH CDR3；Wherein the VL CDR1 includes SEQ ID NO:1；Wherein the VL CDR2 includes SEQ ID NO:2；Wherein the VL CDR3 includes SEQ ID NO:3；Wherein the VH CDR1 includes SEQ ID NO:4；Wherein should VH CDR2 includes SEQ ID NO:5；Wherein the VH CDR3 includes SEQ ID NO:6；Wherein the heavy chain constant region includes SEQ ID NO:28；And wherein the antibody specificity combines the C1s of activation.
The other aspects of the disclosure are related to a kind of immunoconjugates comprising antibody disclosed herein.
The other aspects of the disclosure are related to a kind of nucleotide for encoding antibody disclosed herein or nucleotide set group.
The other aspects of the disclosure are related to inhibiting the method for complement pathway in subject in need, comprising tested to this Person applies the antibody disclosed herein of pharmacy effective dose, immunoconjugates, or nucleotide.
The other aspects of the disclosure are related to the method that the disease or illness of complement-mediated are treated in subject in need, Antibody disclosed herein comprising applying pharmacy effective dose to the subject, immunoconjugates, or nucleotide.
Brief description
Fig. 1 describes the amino acid sequence (SEQ ID NO:10) of humanization VH variant 1 and encodes its nucleotide sequence (SEQ ID NO:11).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 2 describes the amino acid sequence (SEQ ID NO:12) of humanization VH variant 2 and encodes its nucleotide sequence (SEQ ID NO:13).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 3 describes the amino acid sequence (SEQ ID NO:14) of humanization VH variant 3 and encodes its nucleotide sequence (SEQ ID NO:15).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 4 describes the amino acid sequence (SEQ ID NO:16) of humanization VH variant 4 and encodes its nucleotide sequence (SEQ ID NO:17).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 5 describes the amino acid sequence (SEQ ID NO:18) of humanization VH variant 5 and encodes its nucleotide sequence (SEQ ID NO:19).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 6 describes the amino acid sequence (SEQ ID NO:20) of humanization V κ variant 1 and encodes its nucleotide sequence (SEQ ID NO:21).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 7 describes the amino acid sequence (SEQ ID NO:22) of humanization V κ variant 2 and encodes its nucleotide sequence (SEQ ID NO:23).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 8 describes the amino acid sequence (SEQ ID NO:24) of humanization V κ variant 5 and encodes its nucleotide sequence (SEQ ID NO:25).CDR, which is defined with protein sequence number, to be shown according to Kabat numbering.CDR nucleotide and egg White matter sequence is underlined.
Fig. 9 shows C1s (the anti-aC1s of parent mouse anti-activation；Also referred to as TNT005) VH (SEQ ID NO:8) and illustrative Amino acid of differences between humanization VH variant.
Figure 10 shows the amino acid between the anti-aC1s VL of parent mouse (SEQ ID NO:7) and illustrative humanization VL variant Difference.
Figure 11 shows the binding characteristic of the humanization variants of the anti-aC1s of mouse.It shows to the straight of the C1s (" aC1s ") of activation Binding is closed, the competitive binding with the anti-aC1s of 50pM biotinylation (" Biot-005 "), and the number of the inhibition to classic complement approach According to.
Figure 12 shows the binding characteristic of the humanization variants of the anti-aC1s of mouse.Provide the knot of the humanization variants of the anti-aC1s of mouse The affinity data of conjunction.
Figure 13 describes to machin with the anti-aC1s variant (VH3/VK2-Fc- of the humanization of 10mg/kg intravenous administration sub4；Also referred to as TNT020) pharmacokinetics (PK) overview and pharmacodynamics (PD) overview.Data are shown in until 650 small after application When time, % complement pathway (CP) activity (for level standard before applying), the and serum-concentration (μ of antibody applied g/mL)。
Figure 14 describes to machin with the VH3/VK2-Fc-sub of 20mg/kg subcutaneous delivery4PK overview and PD overview.Number According to the time for being shown in 55 days after application, %CP activity (is directed to and applies preceding level standard), and the antibody applied Serum-concentration (μ g/mL).Cover pharmacokinetics (circle) and pharmacodynamics (box) overview.CP=complement pathway.
Figure 15 A-15C provides VH3/VK2-Fc-sub4Amino acid sequence.Figure 15 A provides VH3/VK2-Fc-sub4In deposit Fc-sub4Amino acid sequence.Enhancing the amino acid substitution that FcRn is combined is (Figure 15 A) being underlined.Figure 15 B and 15C provides VH3/VK2-Fc-sub4Heavy chain (Figure 15 B) and light chain (Figure 15 C) amino acid sequence.Variable region CDR is to indicate down (Figure 15 B and 15C) of scribing line, and heavy chain constant region (domain Fc) is overstriking (Figure 15 B).
Figure 16 A-16B provides VH3/VK2-Fc-sub4Overall length weight and light chain amino acid sequence.Signal peptide be overstriking and (Figure 16 A and 16B) being underlined；CDR is (Figure 16 A and 16B) being underlined；And heavy chain constant region (Fc) is to add Thick (Figure 16 A).Enhancing the heavy chain constant region amino acid substitution that FcRn is combined is to indicate double underline (Figure 16 A).
Figure 17 A-17B is that diagram is exposed to the target activity of various concentration and the anti-C1s antibody (side of both inactive C1s Frame) or VH3/VK2-Fc-sub4After (circle), serum complement approach (CP) active (Figure 17 A) and haemolysis (Figure 17 B) in vitro Diagram.
Figure 18 A-18B is the anti-aC1s antibody variants of diagram (VH3/VK2 with wild type IgG4 Fc) and VH3/VK2- Fc-sub4The figure of (with the sequence of heavy chain comprising SEQ ID NO:28) pharmacokinetics (Figure 18 A) and pharmacodynamics (Figure 18 B) overview Show.Negative " medium " control is also shown in Figure 18 B.
Definition
As used in this article, term " complement component C1 s " or " C1s " refer to that a kind of diisopropyl fluorophosphate (DFP) is sensitive Sex pilus serine protease cuts complement component C4 and C2 to start the activation of classic complement approach.People is provided in table 1 The wild-type amino acid sequence (SEQ ID NO:9) of C1s.
Table 1: sequence
Term " antibody " and " immunoglobulin " include the antibody for retaining any isotype of the specific binding to antigen Or immunoglobulin." antibody " includes but is not limited to molecule of the antigen binding and includes at least two weights interconnected by disulfide bond (H) the glycoprotein immunoglobulin or its antigen-binding portion thereof of chain and two light (L) chain.Every H chain includes heavy chain variable region (it is abbreviated herein as VH) and heavy chain constant region.Heavy chain constant region includes three constant domains, CH1, CH2And CH3.Every light chain packet (V is abbreviated herein as containing light chain variable regionL) and constant region of light chain.Constant region of light chain includes a constant domain, CL。VHAnd VL Area can be further subdivided into the denatured region for being referred to as complementary determining region (CDR), be referred to as frame with more conservative The region in area (FR) is spread together.Each VHAnd VLComprising three CDR and four FR, with following order from aminoterminal to carboxyl End arrangement: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.The variable region of weight and light chain is contained and antigen interactions Binding domain.Combination of the constant region energy mediated immunity globulin of antibody to host tissue or the factor, including the various of immune system The first composition (C1q) of cell (such as effector cell) and classical complement system.
For example, term " antibody " includes naturally occurring and non-naturally occurring antibody；Monoclonal and Anti-TNF-α Body；Chimeric and humanized antibody；People or non-human antibody；Fully synthetic antibody；And single-chain antibody.It can be non-by recombination method humanization Human antibody is to reduce it in the intracorporal immunogenicity of people.In the case where explanation is not known, unless otherwise indicated by context, otherwise Term " antibody " further includes the antigen-binding fragment or antigen-binding portion thereof of any foregoing immunoglobulin, and including unit price and two Valence segment or part and single-chain antibody.The antigen-binding fragment of antibody may include that the reservation of antibody combines the target of the antibody Any part of ability.In some embodiments, the antigen-binding fragment of anti-C1s antibody retains the ability for combining C1s.One In a little embodiments, the antigen-binding fragment of antibody includes 1,2,3,4,5 or 6 CDR of the antibody.In some embodiments In, the antigen-binding fragment of antibody includes 1,2,3,4,5,6,7 or 8 frame of 1,2,3,4,5 or 6 CDR and the antibody Area.In some embodiments, the antigen-binding fragment of antibody includes the area VH and/or the area VL of the antibody.
The antibody can (for example) use radioactive isotope, generate the enzyme of detectable product, fluorescin etc. is detectably Label.The antibody can further with other aglucons (such as the member of specific binding pair, such as biotin (biotin-avidin Specific binding pair member)) etc. conjugation.The antibody can also with solid support (including but not limited to polystyrene plate or Pearl) etc. combine.The term further includes monoclonal monomer.As used herein, monoclonal antibody is produced by one group of same cell Raw antibody, all these cells are all to be generated by duplicate cellular replication from single cell.I.e. the clone of cell is only Generate single antibody type.Although hybridoma production technology can be used to generate monoclonal antibody, this field skill can also be used Other production methods known to art personnel (for example originating from the antibody in antibody phage display library).Antibody can be unit price or two Valence.Antibody can be Ig monomer, by 4 polypeptide chains: two heavy chains and two light chains connected by disulfide bond form " Y-Shaped " molecule.
Term " monoclonal antibody " (" mAb ") refers to non-naturally occurring antibody molecule (the i.e. one of individual molecule composition The essentially identical antibody molecule of grade sequence), and it shows the single binding specificity and affinity for defined epitope. MAb is the example of separated antibody.Term " monoclonal antibody " is not limited to the antibody prepared using hybridoma technology.On the contrary, As used herein, monoclonal antibody can be recombinated by hybridoma, transgenosis or other technologies well known by persons skilled in the art It generates.
Term " Humanized immunoglobulin " or " humanized antibody " as used herein refer to immune comprising Different Origin The immunoglobulin of the part of globulin, wherein at least one part include the amino acid sequence of people origin.For example, the humanization Antibody may include the immunoglobulin with essential specificity derived from non-human origins (such as mouse), and be derived from The part of the immunoglobulin sequences (such as chimeric immunoglobulin) of people origin, these parts chemically pass through routine Technology (such as synthesis) combines or using technique for gene engineering (for example, encoding the protein part of the chimeric antibody DNA can obtain expressing to generate continuous polypeptide chain) it is prepared into continuous polypeptide.Another example of Humanized immunoglobulin is Light and/or heavy chain frame containing one or more CDR comprising the antibody derived from non-human origins and derived from human origin The immunoglobulin (such as antibody of the CDR implantation changed with or without frame) of the immunoglobulin chain in area.Some Corresponding ammonia of the largely or entirely amino acid from human immunoglobulin(HIg) in embodiment, except the CDR region of non-human antibody The replacement of base acid.It is some except CDR region in an embodiment of the antibody of humanization form, largely or entirely amino Acid is and some in one or more CDR regions with the amino acid substitution from human immunoglobulin(HIg), largely or entirely ammonia Base acid has not been changed.Term Humanized immunoglobulin further includes chimeric or the single-chain antibody of CDR implantation.See, e.g. Cabilly et al., U.S. Patent number 4,816,567；Cabilly et al., 0,125,023 B1 of european patent number；Boss et al., U.S. Patent number 4,816,397；Boss et al., 0,120,694 B1 of european patent number；Neuberger, M.S. et al., WO 86/ 01533；Neuberger, M.S. et al., 0,194,276 B1 of european patent number；Winter, U.S. Patent number 5,225,539； Winter, 0,239,400 B1 of european patent number；Padlan, E.A. et al., 0,519,596 A1 of European Patent Application No..Also join See, Ladner et al., U.S. Patent number 4,946,778；Huston, U.S. Patent number 5,476,786；And Bird, R.E. et al., Science, 242:423-426 (1988)), about single-chain antibody.Amino acid is added, lacks, is inserted into, replaces or is repaired on a small quantity Decorations are allowed, as long as they do not eliminate the ability of the antibody combination specific antigen.Particularly, one or more frames of the antibody Conserved amino acid in frame area replaces within the scope of this disclosure." humanization " antibody retains the antigentic specificity with original antibodies Similar antigentic specificity.
For example, using synthesis and/or recombinant nucleic acid with prepare encode desired humanization chain gene (such as CDNA) Humanized immunoglobulin is generated.For example, PCR method of mutagenesis can be used to change the DNA of encoding human or humanization chain Sequence (such as DNA profiling from previous humanization variable region) constructs nucleic acid (such as DNA) sequence of encoding humanized variable region It arranges (see, e.g. Kamman, M. et al., Nucl.Acids Res., 17:5404 (1989))；Sato, K. et al., Cancer Research, 53:851-856 (1993)；Daugherty, B.L. et al., Nucleic Acids Res., 19 (9): 2471- 2476(1991)；And Lewis, A.P.and J.S.Crowe, Gene, 101:297-302 (1991)).It is closed using these or other Suitable method can easily generate variant.For example, can mutagenic treatment clone variable region, and may be selected coding have it is desired Specificity variant sequence (for example, being selected from phage library；See, for example, Krebber et al., U.S. Patent number 5, 514,548；Hoogenboom et al., WO 93/06213, open on April 1st, 1993).
The humanization of framework region reduces the antibody and induces people-anti-mouse antibody (HAMA) response risk in human body.It can The method of art-recognized measurement immune response is carried out to monitor the HAMA reaction in particular patient or during clinical test.It can Immunogenicity evaluation is carried out to the patient that applied humanized antibody in the beginning of therapy or during entire application.For example, logical It crosses using method known to those skilled in the art (including Applications of surface plasmon resonance (BIACORE) and/or immobilized enzyme- Linked immunosorbent assay (ELISA) analysis) in blood serum sample of the detection from the patient for humanization therapeutic reagent Antibody reacts to measure the HAMA.In many cases, the anti-C1s antibody of humanization disclosed herein in people experimenter substantially HAMA response is not induced.
Certain amino acid of the selection from people variable framework residues for replacing, this be based on they to CDR conformation and/or It is influenced in conjunction with the possibility of antigen.The non-natural juxtaposition that mouse CDR region and people can be changed framework region can lead to non-natural conformation and limit, and remove It is non-that it is corrected by the certain amino acid residues of substitution, otherwise its loss for leading to binding affinity.
" chimeric antibody " refers to wherein antibody of the variable region derived from a species and constant region derived from another species, example As wherein the variable region is derived from mouse antibodies and the antibody of constant regions derived from human's antibody.
It, can be by the antibody (immunoglobulin) from any invertebrate species based on the amino acid sequence of their constant domains " light chain " be appointed as one in two kinds of significantly different types (referred to as κ and λ).According to the amino of the constant domain of their heavy chains Immunoglobulin can be appointed as different classes of by acid sequence.
There are the immunoglobulins of 5 kinds of primary categories: IgA, IgD, IgE, IgG and IgM, and some in these classifications Subclass (isotype), such as IgG1, IgG2, IgG3, IgG4, IgA and IgA2 can be further divided into.These subclass can be further It is divided into type, such as IgG2a and IgG2b." isotype " refer to by weight chain constant area gene coding antibody isotype or subclass (such as IgM or IgG1).
" antigen " antibody refers to the antibody for specifically binding the antigen.For example, anti-C1s antibody specificity combination C1s.
" antigen-binding portion thereof " (also referred to as " antigen-binding fragment ") of antibody refers to that the one or more of antibody retain specifically Property combine the antigen combined by the entire antibody ability segment.
As used herein, term " affinity " refers to that the balance of the Reversible binding of two kinds of medicaments (such as antibody and antigen) is normal It counts and is expressed as dissociation constant (KD).Affinity is directed to greatly at least 1 times of affinity of unrelated amino acid sequence than antibody, greatly extremely It is 2 times few, greatly at least 3- times, greatly at least 4 times, greatly at least 5 times, greatly at least 6 times, greatly at least 7 times, greatly at least 8 times, greatly at least 9 times, It is at least 10 times big, greatly at least 20 times, greatly at least 30 times, greatly at least 40 times, greatly at least 50 times, greatly at least 60 times, greatly at least 70 times, It is at least 80 times big, greatly at least 90 times, greatly at least 100 times, or big at least 1,000 times, or more.Affinity of the antibody to target protein It can be, for example, about 100 nanomoles (nM) are winged to about 0.1nM, about 100nM to about 1 picomole (pM), or about 100nM to about 1 Mole (fM) or more.As used herein, term " affinity " refers to the compound of two or more medicaments after dilution to solution From resistance.In terms of antibody and/or antigen-binding fragment, term " immunoreactivity " and " preferably in combination with " herein may be used It is used interchangeably.
Term " in conjunction with " refers to the direct joint between two molecules, this is because for example covalently, electrostatic, hydrophobic and ion And/or interaction of hydrogen bond, the interaction including such as salt bridge and water bridge.The anti-C1s antibody specificity of the humanization of the disclosure Epitope in conjugated complement C1s albumen." specific binding " refers to at least about 10-7M or bigger (such as 5x10-7M, 10-8M, 5x10-8M or bigger) affinity combination." non-specific binding " refers to more than about 10-7The affinity of M combines, such as with 10-6M, 10-5M, 10-4The affinity such as M combine.In some embodiments, the activation of the anti-C1s antibody specificity combination C1s and non-live Change two kinds of forms, such as with similar affinity.In certain embodiments, the activation of the anti-C1s antibody specificity combination C1s Form, and the inactive form of C1s is not specifically bound.
As used herein, term " CDR " or " complementary determining region " mean in the variable region of both heavy chain and light chain polypeptide The discontinuous antigen binding site of interior discovery.Kabat et al., J.Biol.Chem.252:6609-6616 (1977) are passed through； Kabat et al., U.S.Dept.of Health and Hunam Services, " Sequences of proteins of Immunological interest " (1991) (also referred to as Kabat 1991 herein)；By Chothia et al., J.Mol.Biol.196:901-917 (1987) (also referred to as Chothia 1987 herein)；With MacCallum et al., J.Mol.Biol.262:732-745 (1996) describes CDR, and wherein this definition includes the amino acid residue when being compared to each other Overlapping or subset.However, being intended to using any definition of the CDR for the antibody or its variant for referring to antibody or implantation defined herein With in the range of the term that uses.Amino acid amino acid as defined in every bibliography of above-mentioned reference including CDR is residual Base is listed in following table 2 as comparing.CDR described in Fig. 1-8 is defined according to Kabat 1991.
Table 2:CDR definition
Kabat1 Chothia2 MacCallum3
VH CDR-1 31-35 26-32 30-35
VH CDR-2 50-65 53-55 47-58
VH CDR-3 95-102 96-101 93-101
VL CDR-1 24-34 26-32 30-36
VL CDR-2 50-56 50-52 46-55
VL CDR-3 89-97 91-96 89-96
1Residue numbering according to the above Kabat et al. nomenclature
2Residue numbering according to the above Chothia et al. nomenclature
3Residue numbering according to the above MacCallum et al. nomenclature
As used herein, term " CDR-L1 ", " CDR-L2 ", and " CDR-L3 " respectively refer in light chain variable region One, the second and the three CDR.As used herein, term " CDR-H1 ", " CDR-H2 ", and " CDR-H3 " respectively refer to weight chain variable The the first, the second and the three CDR in area.As used herein, term " CDR-1 ", " CDR-L2 ", and " CDR-L3 " respectively refer to The first, the second and the three CDR of the variable region of each chain.
As used herein, when for when referring to antibody variable region, term " frame " or " FR " to mean the variable region of antibody All amino acid residues except interior CDR region.Variable framework be usually length between about 100-120 amino acid not Continuous amino acid sequence, but be only intended to reference CDR except those of amino acid.As used herein, term " framework region " means The each domain for the frame opened by CDR points.Light chain variable region (area VL) can have 4 framework regions: FR1, FR2, FR3 and FR4.Phase As, heavy chain variable region (VH) can have 4 framework regions: FR1, FR2, FR3 and FR4.
Term " domain Fc " as used herein or " area Fc " mean functional FcR (such as FcRn) binding partners, unless It states otherwise.The domain Fc is the part in the domain Fc corresponding to natural Ig of polypeptide.The natural domain Fc, which is formed, has same the two of another domain Fc Aggressiveness.In one embodiment, " area Fc ", which refers to, starts from papain shearing site (i.e. in IgG in single Ig heavy chain First residue of heavy chain constant region is set to 114 by the 216th residue) hinge area of positive upstream and end at the C of the antibody The part at end.In some embodiments, the complete domain Fc includes at least hinge domain, the domain CH2 and the domain CH3.According to Ig isotype, The area Fc of Ig constant region may include CH2, the domain CH3 and CH4 and hinge area.In certain embodiments, which may include SEQ ID NO:52 (table 3).In certain embodiments, which includes SEQ ID NO:53 (table 3).In certain embodiments In, which includes SEQ ID NO:28 (table 3).
" separation " antibody is identified from the component of its natural surroundings and separation and/or recycling antibody.Its day The pollution components of right environment are the diagnosis that can interfere antibody or the substance of therapeutical uses, and may include enzyme, hormone and other albumen Matter property or non-proteinaceous solute.In some embodiments, by the antibody purification (1) to more than 90%, it is greater than 95%, Or it is greater than 98%, with the poidometer of the antibody measured by (Lowry) method in labor, it is greater than 99% weight, (2) are enough to lead to Cross the degree that at least 15 residues of N-terminal or central amino acid sequence are obtained using rotation cup sequenator, or (3) to passing through dodecane Base sodium sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) uses Coomassie blue or Silver stain under reduction or non reducing conditions The homogeney of acquisition.Isolated antibody includes the antibody iM situ in recombinant cell, because at least the one of the natural surroundings of the antibody Kind component will not exist.In some cases, pass through the antibody of at least one purification step preparative separation.
The term " polypeptide " being interchangeably used herein, " peptide ", and " albumen " refer to the polymerization of the amino acid of any length Form may include genetically the encode and non-amino acid genetically encoded, chemically or biochemical method modification Or derivatization amino acid, and the polypeptide of the peptide backbone with modification.The term includes fusion protein comprising but be not limited to Fusion protein with heterologous amino acid sequence has heterologous and homologous leader sequences, with or without the end N- methionine The fusion of residue；Immune labeled protein；Deng.Polypeptide, peptide or protein can be naturally occurring or recombination.
As used herein, term " treatment (treatment) ", " treatment (treating) ", " treatment (treat) ", etc. Refer to obtain it is desired pharmacologically and/or effect physiologically.For completely or partially preventing disease or its symptom, The effect can be preventative, and/or just partially or completely cures disease and/or the side effect as caused by the disease and (such as subtracts Less or improve one or more symptoms of the disease) for, it can be therapeutic.As used herein, " treatment ", which is covered, appoints Disease in what treatment mammal, the especially intracorporal disease of people, and include: that (a) prevention is susceptible to suffer from the disease but has not diagnosed The disease occurs in subject with the disease；(b) inhibit the disease, that is, prevent its progress；(c) alleviate the disease, i.e., Cause the recession of the disease.
The term " individual " being interchangeably used herein, " subject ", " host ", and " patient " refer to mammal, Including but not limited to murine (rat, mouse), non-human primate, people, canid, felid, ungulate (such as horse, ox, sheep, pig, goat) etc..These terms further include any animal with complement system, such as mammal, fish Class and some invertebrates.Therefore, these terms include the mammal containing complement system, fish and invertebrate companion Companion animal, agricultural animal, work animal, zoo animal and laboratory animal.
" therapeutically effective amount ", " pharmacy effective dose ", " effective quantity (effective amount) ", or " effective quantity (efficacious amount) " refers to when it is applied to mammal or other subjects for when treating disease, it is sufficient to play The amount of the anticomplement C1s antibody of the effect of the such treatment disease.Being somebody's turn to do " therapeutically effective amount " will be anti-according to anticomplement C1s Body, the age of the disease and its seriousness and subject to be treated, the variation such as weight.
" biological sample " includes the various sample types obtained from individual, and can be used for diagnosing or monitoring measurement.This definition packet Include biological source blood and other fluid samples, Solid Tissue Samples such as biopsy samples or tissue culture or Cell and its offspring as derived from it.This definition further includes the sample that operates in any way after obtaining, for example, by reagent at Reason, dissolves or is enriched with certain components, such as polynucleotides.Term " biological sample " includes clinical sample, and further includes in culture Cell, cell supernatant, cell lysate, serum, blood plasma, biological fluid and tissue sample.Term " biological sample " includes Blood fractions such as urine, saliva, celiolymph, interstitial fluid, ocular fluid, synovia, such as blood plasma and serum etc..Term " biological sample " It further include Solid Tissue Samples, tissue culture sample and cell sample.
As used herein, term " substitution " refers to the difference between given sequence and reference sequences." substitution " is not limited to reality The ad hoc approach of the existing sequence." substitution " can be contrasted with " missing ", and " missing " instruction is relative to reference sequences, and one Or multiple amino acid or nucleotide lacking in given sequence.In both cases, it may be said that given sequence have replace or The amino acid or nucleotide of missing, the source regardless of the sequence.Such as, it may be said that relative to reference sequences, given sequence With in the 100th substitution, even if the given sequence is from the beginning (such as synthetically) and to be not by the prominent of the reference sequences Become creation.In some embodiments, substitution may include the more than one amino acid for replacing single amino acids.
As used herein, term " cross competition (cross compete) " and " cross competition (cross Competition), " refer to the ability an of antibody Yu reference antibody competitive binding target antigen.Any method as known in the art Can be used for determining an antibody whether with reference antibody cross competition.For example, BIAcore is analyzed, ELISA test or streaming are thin Born of the same parents' art can be used for proving the cross competition with existing disclosed antibody.Ability of the inhibition antibody of test antibody in conjunction with people C1s Prove that the test antibody can be with reference antibody competitive binding people C1s.In some embodiments, with reference antibody cross competition In conjunction with antigen (such as people C1s) antibody in conjunction with the reference antibody identical epitope.In some embodiments, and with reference to anti- The antibody of body cross competition combination antigen (such as people C1s) is incorporated near the epitope identified by the reference antibody or neighbouring Epitope.In some embodiments, the antibody of antigen (such as people C1s) is incorporated in by the ginseng in conjunction with reference antibody cross competition Examine the epitope in the distal side of the epitope of antibody identification；However, the combination of the antibody and the distal side epitope is enough to destroy the reference antibody With the binding ability of the antigen.If on the antibody and the antigen with the ammonia with reference antibody interaction on the antigen Base acid is identical or the amino acid residue interaction of overlapping, then an antibody identical epitope in conjunction with reference antibody.
Before further describing the disclosure, it should be understood that the present disclosure is not limited to the specific embodiments, therefore certainly It is changeable.It will also be understood that purpose of the term as used herein merely for description specific embodiment, and be not intended to be limited to, because this Scope of disclosure will be limited only by the claims which follow.
In the case where the range of offer value, it should be understood that unless the context clearly determines otherwise, otherwise in the range Upper and lower bound and range described in this in any other described in or each median between median, until lower limit list / 10th of position, are included within the disclosure.It is smaller that these small range of upper and lower bounds can be independently include this It in range, and is also included within the disclosure, is clearly excluded to be limited by any in the range.It include limit in the range In the case where one or two of fixed, the range for excluding either one or two of restriction that those include is also included within this public affairs In opening.In addition, term " about " is herein for referring to approximatively, roughly, about, or in the zone.When term " about " and numerical value When combined use, the range is modified by extending the up-and-down boundary of the numerical value.Therefore, " about 10-20 " mean " about 10 to About 20 ".In general, term " about " can modify number by the variance (such as 10 percentages) of (higher or lower) upward or downward Value is higher or lower than described value.
Unless otherwise defined, all technical and scientific terms used herein have in the general of disclosure fields The logical identical meaning of the normally understood meaning of technical staff.For example, the Concise Dictionary of Biomedicine And Molecular Biology, Juo, Pei-Show, second edition, 2002, CRC publishing houses；The Dictionary of Cell and Molecular Biology, the 3rd edition, 1999, Academic publishing houses；With the Oxford Dictionary Of Biochemistry And Molecular Biology, revised edition, 2000, Oxford University Press is this field skill Art personnel provide the general dictionary of many terms used in the disclosure.Although similar to method described herein and material or wait Same any method and material can also be used for the practice or test of the disclosure, but now describe preferred method and material.Herein All publications referred to are incorporated herein by mentioning stating with disclosure and description method relevant to the publication of reference and/or material Material.
Unit, prefix and symbol are indicated in the form that its Systeme International de Unites (SI) receives. Digital scope includes the number for defining the range.Unless otherwise defined, amino acid sequence is with amino to carboxyl direction from a left side To right writing.Title provided herein is not the limitation to various aspects of the disclosure, can by reference to the whole instruction and It obtains.Therefore, by the way that the term defined immediately below can be defined more fully with its whole reference book.
It should be noted that in as used herein and the appended claims, singular " one (a) ", " one (an) ", " being somebody's turn to do (the) " includes plural referents, unless the context clearly determines otherwise.Thus, for example, to " the anti-C1s of humanization is anti- Mentioning for body " is stated including a variety of such antibody, and states mentioning for " framework region " including to one or more framework regions and this field skill Its equivalent known to art personnel and suchlike mention are stated.It is further noted that the claims can be drafted to exclude to appoint What optional element.Similarly, this statement is intended as using for example " individually ", " only " and the detailed description with claim elements The removing property term of related analogs, or the antecedent basis for using " negative " to limit.
In addition, "and/or" used herein is considered in two specific features or composition (with or without another In the case where one feature or composition) each of specific disclosure.Therefore, as used in such as " A and/or B " phrase herein Term "and/or" is intended to include " A and B ", " A or B ", " A " (individual), and " B " (individual).Similarly, as example " A, B, And/or term "and/or" used in C " phrase is intended to include each of following aspect: A, B and C；A, B or C；A or C；A or B；B or C；A and C；A and B；B and C；A (individual)；B (individual)；With C (individual).
It should be understood that language "comprising" used herein come from anywhere in describing various aspects, additionally provide with " by ... The other similar aspect that composition " and/or " substantially by ... form " describe.
It should be appreciated that certain features of the disclosure (for the sake of clarity, are retouched in the context of individual embodiment State) offer can also be provided in a single embodiment.On the contrary, the various features of the disclosure are (for simplicity, single The described in the text up and down of embodiment) it can also be provided separately or with the offer of any suitable sub-portfolio.Reality related with the disclosure Apply scheme all combinations specifically include in the disclosure, and it is disclosed herein, as each combination is individually and clear Ground discloses the same.In addition, all sub-portfolios of various embodiments and its element specifically include in the disclosure, and herein Middle disclosure, sub-portfolio as each are independent and clearly disclosed herein.
The publication that is discussed herein only because their disclosures before the submitting day of the application and provide.Herein In any content be not necessarily to be construed as recognizing that the disclosure is had no right by formerly open and prior to such publication.In addition, institute The publication date of offer can be different from practical publication date, can need independently to be confirmed.
Detailed description of the invention
The disclosure provides a kind of antibody of conjugated complement C1s albumen, such as humanized antibody (i.e. anticomplement C1s antibody, Also referred to herein as " anti-C1s antibody ", " C1s antibody ", and " theme antibody ") and a kind of nucleotide comprising encoding such antibody The nucleic acid of sequence.In some respects, which combines activity C1s.In certain embodiments, the anti-C1s is anti- Body does not specifically bind inactive C1s.In some respects, which is humanized antibody.In other aspects, this public affairs The anti-C1s antibody opened has one or more improved pharmaco-kinetic properties, such as improved half-life period, stability, etc..Certain The anti-C1s antibody of aspect, the disclosure can be with subcutaneous administration.The disclosure also provides a kind of antibody comprising the disclosure, such as source of people Change the composition of anti-C1s antibody.The disclosure provides the antibody for generating and using the disclosure, nucleic acid, and the method for composition.This public affairs It opens and the method for the disease or illness for the treatment of complement-mediated is provided, involve the antibody of the application disclosure, such as the anti-C1s of humanization resists Body.
Anticomplement C1s antibody
The disclosure provides anticomplement C1s antibody, such as humanization anticomplement C1s antibody, and the pharmacy comprising such antibody Composition.Complement C1s is a kind of tempting target, because of its upstream in complement cascade and the substrate specificity with close limit Property.Interested in some cases is the antibody for specifically binding the C1s of activated form, such as wherein the antibody is not substantive The C1s of property combination inactive form.
The disclosure provides a kind of anticomplement C1s antibody, such as humanization anticomplement C1s antibody, includes:
A) include following heavy chains: i) including amino acid sequence:
(Q/E)VQL(V/Q)QSGAE(V/L)KKPGASVK(L/V)SC(T/A)ASGFNIKDDYIHWV(K/R) QAPGQGLEWIGRIDPADGHTKYAPKFQVK(V/A)TITADTST(S/N)TAY(L/M)(E/Q)LSSL(R/T)SEDTAVY The area VH of YCARYGYGREVFDYWGQGTTVTVSS (SEQ ID NO:26)；And ii) include and listed ammonia in SEQ ID NO:28 Base acid sequence has the area Fc of at least amino acid sequence of 98% amino acid sequence identity, and wherein amino acid 308 is Leu and ammonia Base acid 314 is Ser；With
B) comprising the light chain of following items: i) include amino acid sequence:
DIVLTQSPDSLAVSLGERATISCKASQSVDYDGDSYMNWYQQK(T/P)GQPPK(I/L)LIYDASNLES The VL of GIPARFSGSGSGTDFTLTISSLE (E/P) EDFA (I/V) YYCQQSNEDPWTFGGGTKVEIK (SEQ ID NO:27) Area；And ii) constant region of light chain.
In some aspects of the disclosure, which includes the heavy chain constant region comprising the area Fc.In some embodiments In, which includes constant region for immunoglobulin, such as human IgG constant region (such as IgG Fc), or its variant.One In a little embodiments, the heavy chain constant region derived from human immunoglobulin of the anti-C1s antibody.In some embodiments, this is anti- The heavy chain constant region of C1s antibody includes IgG4 Fc or its variant.
In some embodiments, the heavy chain Fc of the anti-C1s antibody include with 4 Fc of human IgG (SEQ ID NO:52) or SEQ ID NO:28 is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or the amino acid sequence of at least about 99% sequence identity, wherein amino acid 308 is Leu and amino acid 314 be Ser.In some embodiments, the heavy chain Fc of the anti-C1s antibody include with 4 Fc of human IgG (SEQ ID NO:52) or SEQ ID NO:28 has the amino acid sequence of at least about 98% sequence identity, and wherein amino acid 308 is Leu and amino acid 314 be Ser.In some respects, which includes and the same amino of SEQ ID NO:52 or SEQ ID NO:28 at least 99% Acid sequence, wherein amino acid 308 is Leu and amino acid 314 is Ser.In other aspects, the area Fc of anti-C1s antibody includes SEQ ID NO:28, is substantially made from it, or is made from it.
In some aspects, the anti-C1s antibody of the disclosure have one or more improved pharmacokinetics, such as with wild type Fc Area (SEQ ID NO:52) compare longer half-life period, stability, etc..In certain embodiments, comprising including SEQ ID The anti-C1s antibody in the area heavy chain Fc of NO:28 is with longer than with the area wild type Fc, such as the comparable antibody of 4 Fc of human IgG Half-life period.In other embodiments, the anti-C1s antibody is after subcutaneous administration than having the area wild type Fc, such as 4 Fc of human IgG Comparable antibody it is more stable.Thus, in some respects, the anti-C1s antibody of the disclosure can be with subcutaneous administration.
In some embodiments, the heavy chain Fc of the anti-C1s antibody includes the substitution relative to 4 Fc of human IgG.Some In embodiment, which includes the Fc for having at least about 98% sequence identity with 4 Fc of human IgG.In certain implementations In scheme, which includes Fc, wherein amino acid residue corresponding with the amino acid 1 08 of SEQ ID NO:28 is correct It is proline (such as S241P variant of human IgG 4) when comparison.In certain embodiments, which includes Fc, wherein Amino acid residue corresponding with the amino acid 1 15 of SEQ ID NO:28 is glutamic acid (such as human IgG 4 when correctly comparing L248E variant).In certain embodiments, which includes Fc, wherein right with the amino acid 308 of SEQ ID NO:28 The amino acid residue answered is leucine (such as M428L variant of human IgG 4) in correct compare.In certain embodiments, should Anti- C1s antibody includes Fc, wherein amino acid residue corresponding with the amino acid 314 of SEQ ID NO:28 is silk when correctly comparing Propylhomoserin (such as N434S variant of human IgG 4).
In some embodiments, the anti-C1s antibody include Fc, wherein: (a) with the amino acid 1 08 of SEQ ID NO:28 Corresponding amino acid residue is proline in correct compare；(b) amino acid corresponding with the amino acid 1 15 of SEQ ID NO:28 Residue is glutamic acid in correct compare；(c) amino acid residue corresponding with the amino acid 308 of SEQ ID NO:28 is correctly comparing Clock synchronization is leucine；(d) amino acid residue corresponding with the amino acid 314 of SEQ ID NO:28 is ammonia when correctly comparing Acid；(b), or (e) (a), any combination (c), and (d).In some embodiments, which includes Fc, wherein: (a) amino acid residue corresponding with the amino acid 1 08 of SEQ ID NO:28 is proline when correctly comparing；(b) with SEQ ID The corresponding amino acid residue of amino acid 1 15 of NO:28 is glutamic acid in correct compare；(c) with the amino of SEQ ID NO:28 Sour 308 corresponding amino acid residues are leucines in correct compare；And it is (d) corresponding with the amino acid 314 of SEQ ID NO:28 Amino acid residue be serine in correct compare.
In some embodiments, the Fc of the anti-C1s antibody has bigger than human IgG 4 to Fc receptor, such as FcRn Binding affinity.
In some embodiments, which includes to include SEQ ID NO: The Fc of listed amino acid sequence in 28.In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of the humanization) packet Contain: Fc a) comprising listed amino acid sequence in SEQ ID NO:28；And b) comprising the light chain of people V κ constant region.
Variable region
In some embodiments, the antibody of the disclosure includes heavy chain and light chain, and wherein the heavy chain includes weight chain variable (VH) area and heavy chain constant region, and the light chain includes the area light chain variable (VL)；Wherein the area VL includes to include SEQ ID NO:1's VL complementary determining region (CDR) 1, the VL CDR2 comprising SEQ ID NO:2, and the VL CDR3 comprising SEQ ID NO:3, and wherein The area VH includes the VH CDR1 comprising SEQ ID NO:4, the VH CDR2 comprising SEQ ID NO:5, and includes SEQ ID NO:6 VH CDR3；Wherein the heavy chain constant region includes IgG4 constant region, wherein the heavy chain constant region corresponding to SEQ ID NO:28 Amino acid residue 308 is leucine, and the amino acid residue 314 of the heavy chain constant region corresponding to SEQ ID NO:28 is ammonia Acid；And wherein the antibody specificity combines the C1s of activation.In some embodiments, the heavy chain corresponding to SEQ ID NO:28 The amino acid residue 108 of constant region is proline.In some embodiments, the heavy chain constant region corresponding to SEQ ID NO:28 Amino acid residue 115 be glutamic acid.
In some embodiments, the antibody of the disclosure includes heavy chain and light chain, and wherein the heavy chain includes the area VH and heavy chain Constant region, and the light chain include the area VL；Wherein the area VL includes the VL CDR1 comprising SEQ ID NO:1, includes SEQ ID NO: 2 VL CDR2, and the VL CDR3 comprising SEQ ID NO:3, and wherein the area VH includes the VH comprising SEQ ID NO:4 CDR1, the VH CDR2 comprising SEQ ID NO:5, and wherein the heavy chain constant region includes the VH CDR3 comprising SEQ ID NO:6 SEQ ID NO:28；And wherein the antibody specificity combines the C1s of activation.
In some embodiments, which is human antibody.In some embodiments, which is humanized antibody. In some embodiments, which is chimeric antibody.In some embodiments, which is mouse antibody.
In some embodiments, which is monoclonal antibody.In some embodiments, which is that recombination is anti- Body.In some embodiments, which is synthetic antibody.
In some cases, the cutting to complement component C4 that the anti-C1s antibody of the disclosure inhibits C1s to mediate.Some In embodiment, which inhibits the enzymatic activity of the serine protease domain of C1s.In some cases, the disclosure is anti- The cutting to complement component C2 that C1s antibody inhibits C1s to mediate.In some cases, the anti-C1s antibody of the disclosure inhibits C1s The cutting to C4 and C2 mediated.In some embodiments, C1s/aC1s is cut down in the anti-C1s antibody self-loopa of the disclosure.
In some embodiments, which is humanized antibody.In some cases, the humanization of the disclosure Anti- C1s antibody includes at least one humanization VHFramework region.In some cases, the anti-C1s antibody of the disclosure includes at least one Kind humanization VLFramework region.In some cases, the anti-C1s antibody of the disclosure includes at least one humanization VHFramework region and extremely A kind of few humanization VLFramework region.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes following amino acid sequence VL CDR present in column:
DIVLTQSPASLAVSLGQRATISCKASQSVDYDGDSYMNWYQQKTGQPPKILIYDASNLESGIPARFSG SGSGTDFTLNIHPVEEEDAAIYYCQQSNEDPWTFGGGTKLEIK(SEQ ID NO:7).In some cases, the disclosure The anti-C1s antibody of humanization include VH CDR present in following amino acid sequence:
EVQLQQSGAELVRPGASVKLSCTASGFNIKDDYIHWVKQRPEQGLEWIGRIDPADGHTKYAPKFQVKA TITADTSSNTAYLQLSSLTSEDTAVYYCARYGYGREVFDYWGQGTTLTVSS(SEQ ID NO:8).In some cases In, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes VL CDR and SEQ present in SEQ ID NO:7 VH CDR present in ID NO:8.
In some embodiments, which includes one or more VL CDR present in SEQ ID NO:22. In some embodiments, which includes VL CDR1 present in SEQ ID NO:22.In some embodiments, The anti-C1s antibody includes VL CDR2 present in SEQ ID NO:22.In some embodiments, which includes VL CDR3 present in SEQ ID NO:22.In some embodiments, which includes to deposit in SEQ ID NO:14 One or more VH CDR.In some embodiments, which includes VH present in SEQ ID NO:14 CDR1.In some embodiments, which includes VH CDR2 present in SEQ ID NO:14.In some embodiment party In case, which includes VH CDR3 present in SEQ ID NO:14.
In some embodiments, which includes one or more VL CDR present in SEQ ID NO:30. In some embodiments, which includes VL CDR1 present in SEQ ID NO:30.In some embodiments, The anti-C1s antibody includes VL CDR2 present in SEQ ID NO:30.In some embodiments, which includes VL CDR3 present in SEQ ID NO:30.In some embodiments, which includes to deposit in SEQ ID NO:29 One or more VH CDR.In some embodiments, which includes VH present in SEQ ID NO:29 CDR1.In some embodiments, which includes VH CDR2 present in SEQ ID NO:29.In some embodiment party In case, which includes VH CDR3 present in SEQ ID NO:29.
In certain embodiments, which includes the VL comprising SEQ ID NO:1:KASQSVDYDGDSYMN CDR1(CDR-L1).In some embodiments, which includes the VL comprising SEQ ID NO:33:SQSVDYDGDSY CDR1(CDR-L1).In some embodiments, which includes the VL comprising SEQ ID NO:39:DSYMNWY CDR1(CDR-L1)。
In certain embodiments, which includes the VL CDR2 (CDR- comprising SEQ ID NO:2:DASNLES L2).In some embodiments, which includes the VL CDR2 (CDR-L2) comprising SEQ ID NO:34:DAS.? In some embodiments, which includes the VL CDR2 (CDR-L2) comprising SEQ ID NO:40:ILIYDASNLE.
In certain embodiments, which includes the VL CDR3 comprising SEQ ID NO:3:QQSNEDPWT (CDR-L3).In some embodiments, which includes the VL CDR3 comprising SEQ ID NO:35:SNEDPW (CDR-L3).In some embodiments, which includes the VL CDR3 comprising SEQ ID NO:41:QQSNEDPW (CDR-L3)。
In certain embodiments, which includes the VH CDR1 (CDR- comprising SEQ ID NO:4:DDYIH H1).In some embodiments, which includes the VH CDR1 (CDR- comprising SEQ ID NO:36:GFNIKDD H1).In some embodiments, which includes the VH CDR1 (CDR-H1) comprising SEQ ID NO:42:KDDYIH.
In certain embodiments, which includes the VH comprising SEQ ID NO:5:RIDPADGHTKYAPKFQV CDR2(CDR-H2).In some embodiments, which includes the VH CDR2 comprising SEQ ID NO:37:ADG (CDR-H2).In some embodiments, which includes the VH comprising SEQ ID NO:43:WIGRIDPADGHTK CDR2(CDR-H2)。
In certain embodiments, which includes the VH CDR3 comprising SEQ ID NO:6:YGYGREVFDY (CDR-H3).In some embodiments, which includes the VH CDR3 comprising SEQ ID NO:38:GYGREVFD (CDR-H3).In some embodiments, which includes the VH comprising SEQ ID NO:44:ARYGYGREVFD CDR3(CDR-H3)。
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes to include cdr amino acid Sequence SEQ ID NO:1, SEQ ID NO:2, and the light chain of SEQ ID NO:3 (being CDR-L1, CDR-L2, and CDR-L3 respectively) Variable region.
In some embodiments, the anti-C1s antibody of the disclosure include comprising cdr amino acid sequence SEQ ID NO:4, SEQ ID NO:5, and the heavy chain variable region of SEQ ID NO:6 (being CDR-H1, CDR-H2, and CDR-H3 respectively).
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes to include following sequence The area VH:
(Q/E)VQL(V/Q)QSGAE(V/L)KKPGASVK(L/V)SC(T/A)ASGFNIKDDYIHWV(K/R) QAPGQGLEWIGRIDPADGHTKYAPKFQVK(V/A)TITADTST(S/N)TAY(L/M)(E/Q)LSSL(R/T)SEDTAVY YCARYGYGREVFDYWGQGTTVTVSS(SEQ ID NO:26)。
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes to include the ammonia with table 3 Base acid sequence has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% amino The area VH of the amino acid sequence of acid sequence identity.In certain embodiments, the area VH of the antibody includes selected from by SEQ ID NO:10,12,14,16, and 18 (tables 3) composition group amino acid sequence.In some embodiments, the area the VH packet of the antibody The NO:14 of ID containing SEQ.
Table 3: variable heavy chain variant
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 1 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:10, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VH, wherein amino acid 1 is Glu, ammonia Base acid 5 is Val, and amino acid 11 is Leu, and amino acid 12 is Lys, and amino acid 13 is Lys, and amino acid 20 is Leu, and amino acid 23 is Thr, amino acid 38 are Lys, and amino acid 40 is Ala, and amino acid 42 is Gly, and amino acid 67 is Ala, and amino acid 75 is Thr, ammonia Base acid 76 is Asn, and amino acid 80 is Leu, and amino acid 81 is Gln, and amino acid 83 is Thr, and amino acid 1 09 is Val, wherein ammonia What the number of base acid was described as shown in figure 1.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 1 And SEQ ID NO:10 in listed amino acid sequence the area VH.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 2 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:12, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VH, wherein amino acid 1 is Glu, ammonia Base acid 5 is Val, and amino acid 11 is Val, and amino acid 12 is Lys, and amino acid 13 is Lys, and amino acid 20 is Leu, and amino acid 23 is Thr, amino acid 38 are Lys, and amino acid 40 is Ala, and amino acid 42 is Gly, and amino acid 67 is Ala, and amino acid 75 is Thr, ammonia Base acid 76 is Asn, and amino acid 80 is Leu, and amino acid 81 is Glu, and amino acid 83 is Arg, and amino acid 1 09 is Val, wherein ammonia The number of base acid in Fig. 2 as described.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 2 And SEQ ID NO:12 in listed amino acid sequence the area VH.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 3 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:14, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VH, wherein amino acid 1 is Gln, ammonia Base acid 5 is Val, and amino acid 11 is Val, and amino acid 12 is Lys, and amino acid 13 is Lys, and amino acid 20 is Leu, and amino acid 23 is Thr, amino acid 38 are Lys, and amino acid 40 is Ala, and amino acid 42 is Gly, and amino acid 67 is Val, and amino acid 75 is Thr, ammonia Base acid 76 is Ser, and amino acid 80 is Leu, and amino acid 81 is Glu, and amino acid 83 is Arg, and amino acid 1 09 is Val, wherein ammonia The number of base acid in Fig. 3 as described.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 3 And SEQ ID NO:14 in listed amino acid sequence the area VH.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 4 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:16, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VH, wherein amino acid 1 is Gln, ammonia Base acid 5 is Val, and amino acid 11 is Val, and amino acid 12 is Lys, and amino acid 13 is Lys, and amino acid 20 is Val, and amino acid 23 is Thr, amino acid 38 are Arg, and amino acid 40 is Ala, and amino acid 42 is Gly, and amino acid 67 is Val, and amino acid 75 is Thr, ammonia Base acid 76 is Ser, and amino acid 80 is Met, and amino acid 81 is Glu, and amino acid 83 is Arg, and amino acid 1 09 is Val, wherein ammonia The number of base acid in Fig. 4 as described.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 4 And SEQ ID NO:16 in listed amino acid sequence the area VH.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 5 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:18, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VH, wherein amino acid 1 is Gln, ammonia Base acid 5 is Val, and amino acid 11 is Val, and amino acid 12 is Lys, and amino acid 13 is Lys, and amino acid 20 is Val, and amino acid 23 is Ala, amino acid 38 are Arg, and amino acid 40 is Ala, and amino acid 42 is Gly, and amino acid 67 is Val, and amino acid 75 is Thr, ammonia Base acid 76 is Ser, and amino acid 80 is Met, and amino acid 81 is Glu, and amino acid 83 is Arg, and amino acid 1 09 is Val, wherein ammonia Base acid is numbered as depicted in Figure 5.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 5 And SEQ ID NO:18 in listed amino acid sequence the area VH.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes to include following sequence The area VL:
DIVLTQSPDSLAVSLGERATISCKASQSVDYDGDSYMNWYQQK(T/P)GQPPK(I/L)LIYDASNLES GIPARFSGSGSGTDFTLTISSLE(E/P)EDFA(I/V)YYCQQSNEDPWTFGGGTKVEIK(SEQ ID NO:27)。
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes to include the ammonia with table 4 Base acid sequence has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% amino The area VL of the amino acid sequence of acid sequence identity.In certain embodiments, the area VL of the antibody includes selected from by SEQ ID NO:20,22, and 24 (tables 4) composition group amino acid sequence.In some embodiments, the area VL of the antibody includes SEQ ID NO:22。
Table 4: variable light variant
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 6 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:20, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VL, wherein amino acid 9 is Asp, ammonia Base acid 17 is Glu, and amino acid 40 is Thr, and amino acid 46 is Ile, and amino acid 74 is Thr, and amino acid 76 is Ser, amino acid 77 It is Ser, amino acid 78 is Leu, and amino acid 80 is Glu, and amino acid 83 is Phe, and amino acid 85 is Ile, and amino acid 1 04 is Val, wherein the number of amino acid in Fig. 6 as described.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 6 And SEQ ID NO:20 in listed amino acid sequence the area VL.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 7 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:22, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VL, wherein amino acid 9 is Asp, ammonia Base acid 17 is Glu, and amino acid 40 is Pro, and amino acid 46 is Ile, and amino acid 74 is Thr, and amino acid 76 is Ser, amino acid 77 It is Ser, amino acid 78 is Leu, and amino acid 80 is Pro, and amino acid 83 is Phe, and amino acid 85 is Ile, and amino acid 1 04 is Val, wherein the number of amino acid in Fig. 7 as described.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 7 And SEQ ID NO:22 in listed amino acid sequence the area VL.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure include comprising with retouched in Fig. 8 Listed amino acid sequence has at least 80%, at least 85%, at least 90%, at least 95% in draw and SEQ ID NO:24, until Few 98%, at least 99%, or 100% amino acid sequence identity amino acid sequence the area VL, wherein amino acid 9 is Asp, ammonia Base acid 17 is Glu, and amino acid 40 is Pro, and amino acid 46 is Leu, and amino acid 74 is Thr, and amino acid 76 is Ser, amino acid 77 It is Ser, amino acid 78 is Leu, and amino acid 80 is Pro, and amino acid 83 is Phe, and amino acid 85 is Val, and amino acid 1 04 is Val, wherein the number of amino acid in Fig. 8 as described.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes comprising describing in Fig. 8 And SEQ ID NO:24 in listed amino acid sequence the area VL.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 1 With 1 amino acid sequence of VH variant listed in such as SEQ ID NO:10；And b) describe in Fig. 6 and as listed in SEQ ID NO:20 1 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 1 With 1 amino acid sequence of VH variant listed in such as SEQ ID NO:10；And b) describe in Fig. 7 and as listed in SEQ ID NO:22 2 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 1 With 1 amino acid sequence of VH variant listed in such as SEQ ID NO:10；And b) describe in Fig. 8 and as listed in SEQ ID NO:24 5 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 2 With 2 amino acid sequence of VH variant listed in such as SEQ ID NO:12；And b) describe in Fig. 6 and as listed in SEQ ID NO:20 1 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 2 With 2 amino acid sequence of VH variant listed in such as SEQ ID NO:12；And b) describe in Fig. 7 and as listed in SEQ ID NO:22 2 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 2 With 2 amino acid sequence of VH variant listed in such as SEQ ID NO:12；And b) describe in Fig. 8 and as listed in SEQ ID NO:24 5 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 3 With 3 amino acid sequence of VH variant listed in such as SEQ ID NO:14；And b) describe in Fig. 6 and as listed in SEQ ID NO:20 1 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 3 With 3 amino acid sequence of VH variant listed in such as SEQ ID NO:14；And b) describe in Fig. 7 and as listed in SEQ ID NO:22 2 amino acid sequence of VL variant.In some embodiments, which includes: a) comprising having extremely with SEQ ID NO:14 Few about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or At least about area VH of the amino acid sequence of 99% percentage identity；And b) comprising having at least about with SEQ ID NO:22 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least The area VL of the amino acid sequence of about 99% percentage identity.In some embodiments, which includes: a) including The area VH of the amino acid sequence of SEQ ID NO:14；And b) comprising SEQ ID NO:22 amino acid sequence the area VL.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 3 With 3 amino acid sequence of VH variant listed in such as SEQ ID NO:14；And b) describe in Fig. 8 and as listed in SEQ ID NO:24 5 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 4 With 4 amino acid sequence of VH variant listed in such as SEQ ID NO:16；And b) describe in Fig. 6 and as listed in SEQ ID NO:20 1 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 4 With 4 amino acid sequence of VH variant listed in such as SEQ ID NO:16；And b) describe in Fig. 7 and as listed in SEQ ID NO:22 2 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 4 With 4 amino acid sequence of VH variant listed in such as SEQ ID NO:16；And b) describe in Fig. 8 and as listed in SEQ ID NO:24 5 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 5 With 5 amino acid sequence of VH variant listed in such as SEQ ID NO:18；And b) describe in Fig. 6 and as listed in SEQ ID NO:20 1 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 5 With 5 amino acid sequence of VH variant listed in such as SEQ ID NO:18；And b) describe in Fig. 7 and as listed in SEQ ID NO:22 2 amino acid sequence of VL variant.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) describing in Fig. 5 With 5 amino acid sequence of VH variant listed in such as SEQ ID NO:18；And b) describe in Fig. 8 and as listed in SEQ ID NO:24 5 amino acid sequence of VL variant.
In specific embodiments, which includes the area VH comprising SEQ ID NO:14, includes SEQ ID NO:22's The area VL, and heavy chain constant region；Wherein the heavy chain constant region includes IgG4 constant region, wherein corresponding to the heavy chain of SEQ ID NO:28 The amino acid residue 308 of constant region is leucine, and the amino acid residue 314 of the heavy chain constant region corresponding to SEQ ID NO:28 It is serine；And wherein the antibody specificity combines the C1s of activation.In another embodiment, which includes to include SEQ The area VH of ID NO:14, the area VL comprising SEQ ID NO:22, and the heavy chain constant region comprising SEQ ID NO:28；Wherein this is anti- The C1s of body specific binding activation.
In some embodiments, the anti-C1s antibody include comprising with SEQ ID NO:29 have at least about 80%, at least About 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% is same The heavy chain of the amino acid sequence of one property.As a non-limitative example, in some cases, the antibody of the disclosure (such as people The anti-C1s antibody of sourceization) it include the heavy chain comprising listed amino acid sequence in SEQ ID NO:29.
In some embodiments, the anti-C1s antibody include comprising with SEQ ID NO:30 have at least about 80%, at least About 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% is same The light chain of the amino acid sequence of one property.As a non-limitative example, in some cases, the antibody of the disclosure (such as people The anti-C1s antibody of sourceization) it include the light chain comprising listed amino acid sequence in SEQ ID NO:30.
In some embodiments, which includes the heavy chain comprising SEQ ID NO:29 and includes SEQ ID NO:30's Light chain.
In some embodiments, which includes: a) comprising having at least about 80% with SEQ ID NO:29, At least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The heavy chain of the amino acid sequence of 99% identity；And b) comprising having at least about 80%, at least about 85% with SEQ ID NO:30, At least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or the ammonia of at least about 99% identity The light chain of base acid sequence.In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure includes: a) including SEQ The heavy chain of listed amino acid sequence in ID NO:29；And light chain b) comprising listed amino acid sequence in SEQ ID NO:30.? In some embodiments, in order to generate such antibody, it can be used comprising institute in coding SEQ ID NO:31 and SEQ ID NO:32 The nucleic acid of the nucleotide sequence of column amino acid sequence.
In certain embodiments, which includes constant region of light chain.In some embodiments, the light chain is permanent Determining area includes SEQ ID NO:45
(RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYS LSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC)。
Binding affinity
In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is combined from complement system The complement C1s protein of individual.In some embodiments, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is in conjunction with next From the mammal with complement system, the complement C1s protein of fish, or invertebrate.In some embodiments, this public affairs The antibody (such as the anti-C1s antibody of humanization) opened combines mammal complement C1s protein.In some embodiments, this public affairs The antibody (such as the anti-C1s antibody of humanization) opened combines people's complement C1s protein.In some embodiments, the disclosure is anti- Body (such as the anti-C1s antibody of humanization) combines rat complement C1s protein.In some embodiments, the antibody (example of the disclosure Such as the anti-C1s antibody of humanization) combine the complement C1s protein with the amino acid sequence (SEQ ID NO:9) described in Figure 13. Amino acid sequence SEQ ID NO:9 representative (Homo sapiens) complement C1s protein, with amino acid listed in Figure 13 Sequence.
In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with the dissociation constant no more than 2.5nM (KD) conjugated complement C1s protein.In some embodiments, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is not with K more than 2nMDConjugated complement C1s protein.In some embodiments, (such as the anti-C1s of humanization is anti-for the antibody of the disclosure Body) with the K no more than 1.5nMDConjugated complement C1s protein.In some embodiments, antibody (such as the source of people of the disclosure Change anti-C1s antibody) with the K no more than 1nMDConjugated complement C1s protein.In some embodiments, the antibody (example of the disclosure Such as the anti-C1s antibody of humanization) to be no more than 0.9nM, it is no more than 0.8nM, is no more than 0.7nM, is no more than 0.6nM, is no more than 0.5nM is no more than 0.4nM, is no more than 0.3nM, is no more than 0.2nM, the K no more than 0.1nMDConjugated complement C1s protein.? In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with the K no more than 0.3nMDConjugated complement C1s albumen Matter.In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with the K no more than 0.2nMDConjugated complement C1s Protein.In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with the K no more than 0.1nMDIn conjunction with benefit Body C1s protein.Measurement antibody can be determined by those skilled in the art the method for the combination of C1s protein.
In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is no more than with being no more than 90pM 80pM is no more than 70pM, is no more than 60pM, is no more than 50pM, is no more than 40pM, is no more than 30pM, is no more than 20pM, is no more than 10pM is no more than 9pM, is no more than 8pM, is no more than 7pM, is no more than 6pM, is no more than 5pM, is no more than 4pM, is no more than 3pM, no K more than 2pM, no more than 1pMDConjugated complement C1s protein.
In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with the dissociation constant no more than 2.5nM (KD) combine people's complement C1s protein.In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is not to surpass Cross the dissociation constant (K of 1.5nMD) combine people's complement C1s protein.In some embodiments, antibody (such as the people of the disclosure The anti-C1s antibody of sourceization) with the K no more than 2nMDIn conjunction with people's complement C1s protein.In some cases, the antibody (example of the disclosure Such as the anti-C1s antibody of humanization) with the K no more than 1nMDIn conjunction with people's complement C1s protein.In some cases, the disclosure is anti- Body (such as the anti-C1s antibody of humanization) is no more than 0.8nM to be no more than 0.9nM, is no more than 0.7nM, is no more than 0.6nM, does not surpass 0.5nM is crossed, 0.4nM is no more than, is no more than 0.3nM, is no more than 0.2nM, the K no more than 0.1nMDIn conjunction with people's complement C1s albumen Matter.In some embodiments, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with the K no more than 0.3nMDIn conjunction with people Complement C1s protein.In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with the K no more than 0.2nMD In conjunction with people's complement C1s protein.In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is to be no more than The K of 0.1nMDIn conjunction with people's complement C1s protein.The method for measuring the combination of antibody on human C1s protein can be by this field skill Art personnel determine.In some cases, antibody and people's C1s protein are measured using the binding assay as described in embodiment Between KD。
In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is no more than with being no more than 90pM 80pM is no more than 70pM, is no more than 60pM, is no more than 50pM, is no more than 40pM, is no more than 30pM, is no more than 20pM, is no more than 10pM is no more than 9pM, is no more than 8pM, is no more than 7pM, is no more than 6pM, is no more than 5pM, is no more than 4pM, is no more than 3pM, no K more than 2pM, no more than 1pMDIn conjunction with people's complement C1s protein.
In some cases, the antibody (such as the anti-C1s antibody of humanization) of the disclosure is with 10-8M or less, 5x10-9M or Less, or 10-9Half maximum suppression concentration (IC of M or less50) inhibit classic complement approach.
The antibody (such as the anti-C1s antibody of humanization) of the disclosure can be by complement pathway when being applied to individual in need (CP) activity reduces by 10% to 100% (such as 10% to 15%, 15% to 20%, 20% to 25%, 25% to 30%, 30% To 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90%, or 90% to 100%) Up to 1 day to 1 week, 1 week to 2 weeks, 2 weeks to 4 weeks, 4 weeks to 2 months, or the period more than 2 months.
Such as in some cases, the antibody (such as the anti-C1s antibody of humanization) of the single dose disclosure be applied to it is in need Individual when CP activity can reduce to 10% to 100% (such as 10% to 15%, 15% to 20%, 20% to 25%, 25% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90%, or 90% To 100%) up to 1 day to 1 week, 1 week to 2 weeks, 2 weeks to 4 weeks, 4 weeks to 2 months, or the period more than 2 months.
The antibody (such as the anti-C1s antibody of humanization) of the disclosure can provide effectively when being applied to individual in need by CP Activity reduce by 10% to 100% (such as 10% to 15%, 15% to 20%, 20% to 25%, 25% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90%, or 90% to 100%) up to 1 It to 1 week, 1 week to 2 weeks, 2 weeks to 4 weeks, 4 weeks to 2 months, or the period more than 2 months serum-concentration the humanization Anti- C1s antibody.
Nucleic acid, expression vector and host cell
The disclosure provides the nucleotide sequence comprising anti-C1s antibody (such as the anti-C1s antibody of humanization) disclosed in code book Nucleic acid.In some cases, the nucleic acid of the disclosure includes anti-C1s antibody (such as the anti-C1s antibody of humanization) disclosed in code book The nucleotide sequence in the area VH.In some cases, the nucleic acid of the disclosure includes anti-C1s antibody (such as humanization disclosed in code book Anti- C1s antibody) the area VL nucleotide sequence.In some cases, the nucleic acid of the disclosure includes anti-C1s disclosed in code book anti- The area VH of body (the anti-C1s antibody of such as humanization) and the nucleotide sequence in the area VL.
The nucleotide sequence of anti-C1s antibody (the anti-C1s antibody of such as humanization) disclosed in code book can be operably connected Allow the expression in scheduled target cell (such as through genetic modification with the cell of the antibody of composite coding) should to one or more The controlling element of nucleotide sequence, such as promoter and enhancer.Therefore, in some cases, the disclosure is provided comprising coding The nucleic acid of the nucleotide sequence of the anti-C1s antibody (the anti-C1s antibody of such as humanization) of the disclosure, wherein the nucleotide sequence can operate Ground is connected to one or more controlling elements, such as promoter and/or enhancer.
Suitable promoter and enhancer element is known in the art.Suitable promoter for prokaryotic host cell Including but not limited to phage t7 RNA polymerase promoter；T3 promoter；T5 promoter；λ P promoter；Trp promoter；Lactose Operon promoter；Hybrid promoter, such as lac/tac hybrid promoter, tac/trc hybrid promoter, trp/lac promoter, T7/lac promoter；Trc promoter；Tac promoter etc.；Gpt promoter；AraBAD promoter；The promoter regulated and controled in vivo, it is all Such as ssaG promoter or promoter related (referring to such as U.S. Patent Publication No. 20040131637), pagC promoter (Pulkkinen and Miller, J.Bacteriol., 1991:173 (1): 86-93；Alpuche-Aranda et al., PNAS, 1992；89 (21): 10079-83), nirB promoter (Harborne et al. (1992) Mol.Micro.6:2805-2813) etc. (referring to such as Dunstan et al. (1999) Infect.Immun.67:5133-5141；McKelvie et al. (2004) Vaccine 22:3243-3255；With Chatfield et al. (1992) Biotechnol.10:888-892)；Sigma70 promoter, for example, it is total There is sigma70 promoter (referring to such as GenBank registration number AX798980, AX798961 and AX798183)；Stationary phase starting Son, such as dps promoter, spv promoter etc.；Promoter derived from pathogenicity island SPI-2 (see, e.g. WO96/17951)； ActA promoter (see, e.g. Shetron-Rama et al. (2002) Infect.Immun.70:1087-1096)；RpsM starting Sub (see, e.g. Valdivia and Falkow (1996) .Mol.Microbiol.22:367)；Tet promoter (see, e.g. Hillen, W. and Wissmann, A. (1989) In Saenger, W. and Heinemann, U. (eds), Topics in Molecular and Structural Biology, Protein-Nucleic Acid Interaction.Macmillan, London, Britain, the 10th phase, the 143-162 pages)；SP6 promoter is (see, e.g. Melton et al. (1984) Nucl.Acids Res.12:7035)；Deng.The suitable strong promoter of prokaryotes for such as Escherichia coli (Escherichia coli) Including but not limited to Trc, Tac, T5, T7 and Pλ.The unrestricted example of operon for bacterial host cell includes cream (when contacting lactose, LacI aporepressor changes conformation to sugared promoter operon, so that preventing the LacI aporepressor from combining should Operon), (when being complexed with tryptophan, TrpR aporepressor has the structure in conjunction with the operon to trp promoter operon As；In the case where lacking tryptophan, which has the conformation for not combining the operon) and tac promoter behaviour Vertical son (see, e.g. deBoer et al. (1983) Proc.Natl.Acad.Sci.U.S.A.80:21-25).
In some embodiments, such as the expression in yeast cells, suitable promoter is constitutive promoter, Such as ADH1 promoter, PGK1 promoter, ENO promoter, PYK1 promoter etc.；Or regulation type promoter, such as GAL1 starting Son, GAL10 promoter, ADH2 promoter, PHO5 promoter, CUP1 promoter, GAL7 promoter, MET25 promoter, MET3 are opened Mover, CYC1 promoter, HIS3 promoter, ADH1 promoter, PGK promoter, GAPDH promoter, ADC1 promoter, TRP1 are opened Mover, URA3 promoter, LEU2 promoter, ENO promoter, TP1 promoter and AOX1 are (as being used for pichia (Pichia))。
For the expression in eukaryocyte, suitable promoter includes but is not limited to light and/or heavy chain immunoglobulin Gene promoter and enhancer element；Cytomegalovirus early promoter at once；Herpes simplex thymidine kinase promoter； Early and late SV40 promoter；The promoter being present in the long terminal repeats of retrovirus；Mouse metallothionein White-I promoter；With various tissue-specific promoters known in the art.Composing type mammalian promoter includes but unlimited In the promoter for being directed to following gene: hypoxanthine phosphoribosyltransferase (HPRT), adenosine deaminase, pyruvate kinase, β flesh Filamentous actin promoter and other constitutive promoters.The virus acted on to the additional illustrative composition in eukaryocyte Promoter includes, for example, cytomegalovirus (CMV) is come from, simian virus (such as SV40), Papillomavirus, adenovirus, people Immunodeficiency virus (HIV), Rous sarcoma virus cytomegalovirus, the long terminal repeats of moloney leukemia virus (LTR) and the thymidine kinase promoter of the promoter of other retrovirus and herpes simplex virus.Other composing types open Mover is known to persons of ordinary skill in the art.The promoter that can be used as the expressed sequence of the disclosure further includes induction type Promoter.Inducible promoter is expressed in the presence of induction agent.For example, induction should in the presence of certain metal ions Metallothionein promoter is to start transcription and translation.Other inducible promoters are known to persons of ordinary skill in the art.
Select suitable carrier and promoter completely within ordinary skill level.
The nucleic acid of nucleotide sequence comprising anti-C1s antibody disclosed in code book (such as the anti-C1s antibody of humanization) can be deposited It is in any expression vector and/or cloning vector known in the art.As used in this article, expression vector refers to containing transcription Any nucleic acid construct of element necessary to coded sequence with translation insertion contains in the case where rna virus vector, Any nucleic acid construct of element necessary to replicating and translate when it being transferred in suitable host cell.Expression vector can wrap Include plasmid, phasmid, virus and its derivative.
The some aspects of the disclosure are provided comprising including anti-C1s antibody disclosed in code book (the anti-C1s antibody of such as humanization) Nucleotide sequence nucleic acid recombinant vector, wherein the recombinant vector is cloning vector.The some aspects of the disclosure provide packet The recombinant vector of nucleic acid containing the nucleotide sequence comprising anti-C1s antibody disclosed in code book (the anti-C1s antibody of such as humanization), In the recombinant vector be expression vector, for example, wherein the nucleotide sequence be operably coupled to it is suitable in the expression vector Regulating and controlling sequence to ensure the expression of the antibody of the coding.In the case where theme antibody includes two sseparated polypeptides, coding The nucleic acid of two polypeptides can be cloned in identical or separated carrier, to form one or more recombinant vectors.Recombination carries Body may include selected marker, replication orgin, and providing the duplication and/or maintenance of the recombinant vector (such as recombinant expression carrier) Other feature.
Largely suitable carrier and promoter are known to the skilled in the art；It is many commercially available for generating master Inscribe recombinant vector.Following carrier is provided by example.Bacterium: pBs, phagescript, PsiX174, pBluescript SK, PBs KS, pNH8a, pNH16a, pNH18a, pNH46a (Stratagene, La Jolla, Calif., USA)；PTrc99A, PKK223-3, pKK233-3, pDR540 and pRIT5 (Pharmacia, Uppsala, Sweden).Eucaryote: pWLneo, PSV2cat, pOG44, PXR1, pSG (Stratagene) pSVK3, pBPV, pMSG and pSVL (Pharmacia).
Expression vector usually has the convenient restriction site near promoter sequence, to provide encoding heterologous egg The insertion of white nucleic acid sequence.The effective as selective in expressive host may be present to mark.Suitable expression vector includes but not It is limited to viral vectors.The example of viral vectors includes but is not limited to based on following viral vectors: vaccinia virus；Polio Virus；Adenovirus (see, e.g. Li et al. people, Invest Opthalmol Vis Sci 35:2543-2549,1994；Borras Et al., Gene Ther 6:515-524,1999；Li and Davidson, PNAS 92:7700-7704,1995；Sakamoto etc. People, H Gene Ther 5:1088-1097,1999；WO 94/12649, WO 93/03769；WO 93/19191；WO 94/ 28938；WO 95/11984 and WO 95/00655)；Adeno-associated virus is (referring to such as Ali et al., Hum Gene Ther 9:81- 86,1998, Flannery et al., PNAS 94:6916-6921,1997；Bennett et al., Invest Opthalmol Vis Sci 38:2857-2863,1997；Jomary et al., Gene Ther 4:683-690,1997, Rolling et al., Hum Gene Ther 10:641-648,1999；Ali et al., Hum Mol Genet 5:591-594,1996；The WO of Srivastava 93/09239, Samulski et al., J.Vir. (1989) 63:3822-3828；Mendelson et al., Virol. (1988) 166: 154-165；With Flotte et al., PNAS (1993) 90:10613-10617)；SV40；Herpes simplex virus；Retrovirus Carrier (such as murine leukemia virus, spleen necrosis virus, and (such as Rous sarcoma virus breathes out dimension meat derived from retrovirus Tumor virus, avian leukosis virus, human immunodeficiency virus (see, e.g. Miyoshi et al., PNAS 94:10319-23, 1997；Takahashi et al., J Virol 73:7812-7816,1999)), Myeloproliferative Sarcoma virus and mammary tumor virus Carrier)；Deng.
In some embodiments, which is viral vectors.Viral vectors includes but is not limited to from lower influenza virus Nucleic acid sequence: retrovirus, such as Moloney Murine Leukemia virus breathe out dimension rat meat tumor mouse, MuMTV and labor This sarcoma virus；Adenovirus, adeno-associated virus；SV40 type virus；Polyomavirus；Epstein epstein-Barr virus；Papilloma Virus；Herpesviral；Vaccinia virus；Poliovirus；And RNA virus, such as retrovirus.It can be easily with this Other carriers known in field.Eukaryotic viral of certain viral vectors based on non-cell pathogenicity, wherein dispensable gene has been Replaced with interested gene.The virus of non-cell pathogenicity includes retrovirus, and life cycle includes by genome disease Provirus is then integrated into host cell DNA by malicious RNA reverse transcription at DNA.Retrovirus has been approved by for mankind's base Because in therapeutic test.The most useful is that replication defective (can instruct the synthesis of desired protein, but cannot manufacture infection Property particle) those of retrovirus.The retrovirus expression vector of such genetic change has high efficiency transduction base in vivo The general service of cause.For generating the standard scheme of the retrovirus of replication defective (the following steps are included: exogenous genetic material It is incorporated to plasmid, the cell line packed with plasmid transfection generates recombinant retrovirus by package cell line, from tissue culture medium (TCM) Middle collection virion, and with the infestation with virus particles target cell) in Kriegler, M., Gene Transfer and Expression, A Laboratory Manual, W.H.Freeman Co., New York (1990) and Murry, E.J., Methods in Molecular Biology, the 7th phase mention in Humana Press, Inc., Cliffton, N.J. (1991) For.
In an embodiment, which is adeno-associated virus, double-stranded DNA virus.The adeno-associated virus can be built into multiple Deficiency processed and large-scale cell type and type can be infected.It further has following advantage: for example hot and lipid is molten Agent stability；High transduction frequencies in different lineages, including hematopoietic cell；Inhibit with superinfection is lacked to allow multiple Transduction.It is reported that the adeno-associated virus being integrated into people's cell with site-specific fashion, to farthest drop The changeability of the gene expression characteristics of the insertion of a possibility that being inserted into mutagenesis and retroviral infection.In addition, not selecting In the case where selecting pressure, wild type adeno-associated virus infection has then been carried out in tissue cultures and has been passed on more than 100 times, this meaning Adeno-associated virus genome conformity be metastable event.The adeno-associated virus can also be acted as in a manner of extrachromosomal With.
In another embodiment, which operated to carry and encode anti-C1s as disclosed herein The adeno-associated virus (AAV) of the polynucleotides of antibody.The conventional method for obtaining recombination AAV (rAAV) is disclosed.See, e.g., USP 8,734,809,2013/0195801 and references cited therein.In some embodiments, rAAV carrier packet (ITR) and interested transgenosis (such as FIX polynucleotide sequence of optimization) are repeated containing one or more AAV opposing ends. In certain embodiments, the method for preparing rAAV is related to cultivating desired host cell, which contains coding AAV The nucleic acid sequence of capsid protein or its segment；Functional rep gene；AAV opposing end repeats (ITR) and interested transgenosis The rAAV carrier of composition；With enough miscellaneous functions to allow to recombinate AAV carrier package into AAV capsid protein.For reality The material and method and relative program for applying the above are in (for example) USP 8,734,809,2013/0195801, PCT/ US1997/015692, PCT/US2002/033692, PCT/US2002/033630, WO2007/148971, WO00/20561, It is disclosed in WO03/042361 and WO2007/04670.
One or more different AAV carrier sequences derived from substantially any serotype can be used according to the disclosure.It is special The selection for determining AAV carrier sequence will be guided by known parameters, such as taxis interested, required carrier output etc..In general, should AAV serotype, with the genome sequence of significant homology, provides relevant genetic function group on amino acid and nucleic acid level, It generates virion (these virion are relevant), and similarly replicates and assemble.For the base of the various AAV serotypes Because the summary of group sequence and genome similarity is see, for example, GenBank registration number U89790；GenBank registration number J01901； GenBank registration number AF043303；GenBank registration number AF085716；Chlorini et al. (1997, J.Vir.71:6823- 33)；Srivastava et al. (1983, J.Vir.45:555-64)；Chlorini et al. (1999, J.Vir.73:1309- 1319)；Rutledge et al. (1998, J.Vir.72:309-319)；With Wu et al. (2000, J.Vir.74:8635-47).AAV Serotype 1,2,3,4 and 5 is the illustrative source of AAV nucleotide sequence, the context for the disclosure.AAV6, AAV7, AAV8 Or AAV9 or the AAV sample particle developed recently obtained by such as capsid shuffling technology and AAV capsid library or from design recently, Exploitation or the ITR's to evolve are also applied for certain published applications.Referring to Dalkara, D et al. (2013), Sci Transl.Med.5(189):189ra76；Kotterman,MA Nat.Rev.Genet.(2014)15(7):455.
In other embodiments, which is derived from slow virus.In certain embodiments, which is that can infect The carrier of the recombinant slow virus of non-dividing cell.
The lentiviral gene group and the proviral DNA usually have the 3 kinds of genes found in retrovirus: gag, Pol and env, flank are that two long end repeats (LTR) sequence.The gag gene encodes internal structure (matrix, capsid and core Capsid) albumen；The archaeal dna polymerase (reverse transcriptase) of pol gene coding RNA guidance, protease and integrase；And the env base Because encoding viral envelope glycoprotein.The 5' and 3'LTR's is used to promote transcription and the polyadenylation of virion RNA.It should LTR contains all other cis acting sequence necessary to virus replication.Slow virus has additional gene, including vif, vpr, Tat, rev, vpu, nef and vpx (in HIV-l, HIV-2 and/or SIV).
With the 5'LTR it is neighbouring be the reverse transcription genome (tRNA primer binding site) and the effective capsidation of viral RNA At sequence necessary to particle (site Psi).If capsidation (or retrovirus RNA is made to be packaged into infectious virus particle) Necessary sequence lacks in viral genome, then the cis- defect prevents the capsidation of geneome RNA.
However, gained mutant still is able to instruct the synthesis of all virion proteins.The disclosure, which provides, to be generated and can feel Contaminate the method for the recombinant slow virus of non-dividing cell, this method include with two or more carrier pack functions (i.e. gag, pol with Env and rev and tat) the suitable host cell of carrier transfection.As disclosed below, lack functionality tat gene Carrier is desired for certain applications.Thus, for example, first vector can provide the core of coding virus gag and virus pol Acid, and another carrier can provide the nucleic acid of coding virus env, to generate the cell of packaging.Carrier (this of heterologous gene will be provided Text is accredited as transfer vector) it imports this incasing cells and generates release and carry the infectious viral particle of interested foreign gene Production cell.
According to the above structure of carrier and foreign gene, which can provide coding peplos (env) gene Nucleic acid.The env gene can be derived from substantially any suitable virus, including retrovirus.In some embodiments, should Env albumen is double preferendum envelope proteins, allows the cell of transduction human and other species.
The example of env gene derived from retrovirus includes but is not limited to: Moloney Murine Leukemia virus (MoMuLV Or MMLV), Harvey murine sarcoma virus (HaMuSV or HSV), MuMTV (MuMTV or MMTV), gibbon ape leulcemia Viral (GaLV or GALV), human immunodeficiency virus (HIV) and Rous sarcoma virus (RSV).Other env bases also can be used Cause, such as vesicular stomatitis virus (VSV) Protein G (VSV G), the env gene of hepatitis virus and influenza virus.
It is operationally associated with regulating and controlling sequence described at other herein for providing the carrier of viral env nucleic acid sequence.
In certain embodiments, which includes slow virus carrier, wherein HIV virulence gene env, vif, vpr are deleted, Ability of the vpu and nef without damaging the carrier transduction non-dividing cell.
In some embodiments, which includes the slow virus carrier of the missing in the area U3 comprising 3'LTR.The area U3 lacks Mistake can be missing or excalation completely.
In some embodiments, the disclosure of the nucleotide sequence comprising encoding anti-C1s antibody as described herein can be made Slow virus carrier be transfected into (a) include gag, pol or gag and pol gene first nucleotide sequence and (b) include In the cell of second nucleotide sequence of heterologous env gene；Wherein the slow virus carrier lacks functional tat gene.Other In embodiment, the cell further is transfected with the 4th nucleotide sequence comprising rev gene.In certain embodiments, should Slow virus carrier lacks selected from vif, vpr, vpu, vpx and nef, or combinations thereof functioning gene.
In certain embodiments, slow virus carrier includes the nucleotide sequence of one or more encoding gag albumen, Rev- Response element, the poly- purine sequence (cPPT) in center, or any combination thereof.
The slow virus is disclosed in WO9931251, W09712622, W09817815, W09817816 and WO9818934 The example of carrier, is by reference incorporated herein in its entirety.
Other carriers include plasmid vector.Plasmid vector description extensively in the art, and be those skilled in the art It is known.See, e.g. Sambrook et al., Molecular Cloning:A Laboratory Manual, second edition, cold spring Publishing house, Cold Spring Harbor Laboratory, 1989.In recent years, it has been found that plasmid vector is particularly conducive to gene delivery to intracorporal cell, because It cannot replicate and be integrated into host genome in host genome for them.However, having compatible with the host cell The gene expression peptide that these plasmids of promoter can operationally be encoded out of this plasmid.Can be obtained from commercial supplier one Common plasmids include pBR322, pUC18, pUC19 a bit, various pcDNA plasmids, pRC/CMV, various pCMV plasmids, pSV40, and pBlueScript.The additional examples of specific plasmids include pcDNA3.1, catalog number (Cat.No.) V79020；PcDNA3.1/hygro, catalogue Number V87020；PcDNA4/myc-His, catalog number (Cat.No.) V86320；And pBudCE4.1, catalog number (Cat.No.) V53220 all are from Invitrogen(Carlsbad,CA.).Other plasmids are well known to those of ordinary skill in the art.In addition, the standard of can be used Protocols in Molecular Biology custom design plasmid is to remove and/or add the specific fragment of DNA.
Host cell
The disclosure provides (such as external thin with the host cell of the separated genetic modification of theme nucleic acid genetic modification Born of the same parents).In some embodiments, the theme host cell of separated genetic modification can produce theme antibody.Such cell claims For " recombinant cell " or " host cell of genetic modification ".The host cell of the genetic modification of the disclosure includes comprising code book public affairs The nucleic acid of the nucleotide sequence for the anti-C1s antibody (the anti-C1s antibody of such as humanization) opened.
Suitable host cell includes eukaryotic host cell, such as mammalian cell, insect host cell, and yeast is thin Born of the same parents；And prokaryotic cell, such as bacterial cell.Calcium phosphate precipitation, the transfection that deae dextran mediates, liposome can (for example) be passed through The transfection of mediation, electrotransformation or the realization of other known method import theme nucleic acid in the host cell.
Suitable mammalian cell includes primary cell and immortalized cell line.Suitable mammal cell line includes Human cell line, non-human primate's cell line, rodent (such as mouse, rat) cell line etc..Suitable mammal is thin Born of the same parents system includes but is not limited to HeLa cell (such as American Type Tissue Culture institute (ATCC) number CCL-2), Chinese hamster ovary celI (example Such as ATCC number CRL9618, CCL61, CRL9096), 293 cells (such as ATCC number CRL-1573), Vero cell, NIH 3T3 cell (such as ATCC number CRL-1658), Huh-7 cell, bhk cell (such as ATCC number CCL10), PC12 cell (ATCC number CRL1721), COS cell, COS-7 cell (ATCC number CRL1651), CVI (MK cells system), RAT1 is thin Born of the same parents, mouse Lcell (ATCC number CCLI.3), human embryo kidney (HEK) (HEK) cell (ATCC number CRL1573), HLHepG2 cell etc.. In some cases, which is HEK cell.In certain embodiments, which is 293 cell of HEK.In some cases In, which is Chinese hamster ovary celI, such as CHO-K1 cell (ATCC number CCL-61), CHO-M cell, CHO-DG44 cell (ATCC Number PTA-3356), DUXB11 (Chinese hamster ovary line, DHFR minus), (Chinese hamster is at fiber finer by R1610 Born of the same parents), BALBC/3T3 (l cell), HAK (Hamster kidney cell line), SP2/O (mouse myeloma), P3x63- Ag3.653 (mouse myeloma), BFA-1c1BPT (bovine aortic endothelial cells), RAJI (human lymphocyte),NS0, CAP, BHK21 etc..In some embodiments, which is COS cell.In some embodiments, the host cell It is 293 cells.In some embodiments, which is Chinese hamster ovary celI.
Suitable yeast cells includes but is not limited to pichia pastoris yeast (Pichia pastoris), and Finland finishes red ferment Female (Pichia finlandica) is liked trehalose Pichia pastoris (Pichia trehalophila), Pichia kudriavezii (Pichia koclamae), Pichia membranaefaciens (Pichia membranaefaciens) have and embrace Pichia pastoris (Pichia Opuntiae), heat-resisting Pichia pastoris (Pichia thermotolerans), willow Pichia pastoris (Pichia salictaria), Pichia pastoris (Pichia guercuum), Pi Jiepu Pichia pastoris (Pichia pijperi), pichia stipitis (Pichia stiptis), thermophilic pichia methanolica (Pichia methanolica), pichia (Pichia sp.) are made Brewer yeast (Saccharomyces cerevisiae), saccharomyces (Saccharomyces sp.), Hansenula polymorpha (Hansenula polymorpha), Kluyveromyces (Kluyveromyces sp.), Kluyveromyces Lactis ties up this yeast (Kluyveromyces lactis), Candida albicans (Candida albicans), aspergillus nidulans (Aspergillus Nidulans), aspergillus niger (Aspergillus niger), aspergillus oryzae (Aspergillus oryzae), trichoderma reesei (Trichoderma reesei), lucknow gold pityrosporion ovale (Chrysosporium lucknowense), Fusarium (Fusarium sp.), cereal cutting insert bacterium (Fusarium gramineum), Fusarium venenatum, Neuraspora crassa (Neurospora crassa), Chlamydomonas reinhardtii (Chlamydomonas reinhardtii) etc..In some embodiments, should Host cell is saccharomyces.In some embodiments, which is pichia.
Suitable prokaryotic cell includes but is not limited to Escherichia coli, bacillus (Bacillus) (such as bacillus subtilis (B.subtilis)), any one of kinds of experiments room bacterial strain such as genus lactubacillus (Lactobacillus sp.).See, e.g. Carrier et al. (1992) J.Immunol.148:1176-1181；U.S. Patent number 6,447,784；With Sizemore et al. (1995)Science 270:299-302.In general, the laboratory strains are non-pathogenics.In some embodiments, the place Chief cell is Escherichia coli.In some embodiments, which is bacillus subtilis.
The separated nucleic acid molecules of the disclosure can be made to lead by various technology completions well-known to those skilled in the art Enter in the host cell.These technologies include but is not limited to transfect (including electrophoresis and electroporation), protoplast fusion, calcium phosphate Precipitating, cell are merged with coating DNA, the infection of microinjection and intact virus.Referring to Ridgway, A.A.G. " Mammalian Expression Vectors " the 24.2nd chapter, the 470-472 pages carrier, Rodriguez and Denhardt is compiled It collects (Butterworths, Boston, Mass.1988).Most preferably, import plasmid in the host via electroporation.It should Transformed cell is grown under conditions of being suitable for and generating light chain and heavy chain, and should for weight and/or light chain protein synthesis measurement Transformed cell.Exemplary assay techniques include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), Or fluorescence activated cell sorts analysis (FACS), immunohistochemistry etc..
The host cell of separated nucleic acid molecules comprising the disclosure is grown in suitable growth medium.As herein Used, term " suitable growth medium " means the culture medium of the nutrient needed for growing containing cell.Cell grows institute The nutrient needed may include carbon source, nitrogen source, it is necessary to amino acid, vitamin, minerals and growth factor.Optionally, the culture medium One or more selective factor Bs can be contained.Optionally, which can contain calf serum or fetal calf serum (FCS).At one In embodiment, the culture medium is essentially free of IgG.The growth medium for example, by medicament selection or must usually will be sought Support element defect and select the cell containing DNA construct, the essential nutrients by the DNA construct selected marker supplement or With the DNA construct cotransfection.Culture medium of the mammalian cell through cultivating usually commercially available containing serum or serum-free Growth in (such as MEM, DMEM, DMEM/F12).In one embodiment, the culture medium be CDoptiCHO (Invitrogen, Carlsbad,CA.).In another embodiment, which is CD17 (Invitrogen, Carlsbad, CA.).Selection Culture medium suitable for specific cell line used is in the level of those of ordinary skill in the art.
Pharmaceutical compositions
The disclosure provides composition, and the composition includes the anti-C1s antibody (the anti-C1s antibody of such as humanization) comprising the disclosure Pharmaceutical compositions.In general, the anti-C1s that pharmaceutical compositions (referred to herein as preparaton) include a effective amount of disclosure resists Body (the anti-C1s antibody of such as humanization)." effective quantity " means the dosage for being enough to generate desired result, such as reduces and be situated between with complement The relevant ill symptoms of disease or illness led, improve the disease of complement-mediated or the symptom of illness, slow down the disease of complement-mediated Disease or the progress of illness etc..In general, compared with the control, which at least mitigates the disease of complement-mediated or the disease of illness Shape.In some embodiments, the anti-C1s antibody of the disclosure (such as the anti-C1s antibody of humanization) is prepared and/or is modified So that the antibody can pass through blood-brain barrier.In some embodiments, anti-C1s antibody (such as the anti-C1s of humanization of the disclosure Antibody) it is delivered in a manner of such to avoid blood-brain barrier.In some embodiments, the disclosure anti-C1s antibody (such as The anti-C1s antibody of humanization) it is prepared with the medicament across blood-brain barrier is promoted.In some embodiments, the disclosure is anti- C1s antibody (such as the anti-C1s antibody of humanization) is melted directly or through connector with the compound across blood-brain barrier is promoted It closes.
Preparaton
The pharmaceutical compositions can be formulated by bolus infusion for parenteral administration (it is i.e. intravenous, subcutaneously or muscle It is interior).Preparaton for injection can be presented in the form of unit dose, such as in the ampoule of preservative with addition and more In dose container.The composition can take the suspension in such as oiliness or aqueous vehicles, the form of solution or lotion, and Containing formulatory's medicament, such as suspending agent, stabilizer and/or dispersing agent.Alternatively, which can be the shape of powder Formula, for being formed together with suitable medium (such as without heat source water).
In the subject method, the anti-C1s antibody of any convenient means application disclosure can be used, and (such as humanization is anti- C1s antibody) to the host, desired therapeutic effect or diagnosis effect can be generated.Therefore, medicament incorporation can be made various For treating application in preparaton.More specifically, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure can by with Suitable pharmaceutically acceptable carrier, pharmaceutically acceptable diluent or other pharmaceutically acceptable excipient compositions are configured to Pharmaceutical compositions, and it can be configured to solid, semisolid, the preparation of liquid or gas form, for example (,) tablet, capsule, powder, Grain, ointment, solution, suppository, injection, inhalant and aerosol.In some embodiments, pharmaceutical compositions include the disclosure Anti- C1s antibody (such as the anti-C1s antibody of humanization) and pharmaceutically acceptable excipient.
In pharmaceutical dosage form, anti-C1s antibody (such as the people of the disclosure can be applied in the form of its pharmaceutically acceptable salt The anti-C1s antibody of sourceization) or they also can be used alone or be suitably associated with and combine with other pharmaceutically active compounds and make With.Following method and excipient are merely exemplary and restrictive by no means.
For oral preparation, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure can be used alone or with it is suitable Additive be applied in combination to prepare tablet, powder, granule or capsule, it is sweet for example, combined with conventional additives, such as lactose Reveal alcohol, cornstarch or dehydrated potato powder；It is combined with adhesive, such as avicel cellulose, cellulose derivative, Arabic gum, it is beautiful Rice starch or gelatin；It is combined with disintegrating agent, for example, cornstarch, dehydrated potato powder or sodium carboxymethylcellulose；With lubricant group It closes, such as talcum powder or magnesium stearate；And it if desired, can be with diluent, buffer, wetting agent, preservative and flavoring agent combination.
The anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure can be made to be configured to the preparation for injection, by It is dissolved in aqueous or non-aqueous solvent, suspends or emulsify the antibody, for example, vegetable oil or other similar oil, propylene glycol, synthetic fat Fatty acid glyceride, the organic ester (such as ethyl oleate) of injectable, the ester or propylene glycol of higher fatty acids；If desired, available normal Advise additive, such as solubilizer, isotonic agent, suspending agent, emulsifier, stabilizer and preservative.Parenteral medium includes chlorination Sodium solution, woods grignard glucose, dextrose and sodium chloride, Lactated Ringer'S Solution or fixing oil.Intravenous vehicles include liquid And nutritional supplement, electrolyte replenisher (such as electrolyte replenisher based on woods grignard glucose) etc..In addition, the disclosure Pharmaceutical compositions may include other medicament, such as dopamine or psychopharmacological agents, this depends on the pharmaceutical compositions Desired use.
By make the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of the purity with desired level with Optional physiologically acceptable carrier, other excipient, stabilizer, surfactant, buffer and/or tonicity agent mixing are to make The pharmaceutical compositions of the standby anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure.Acceptable carrier, Qi Tafu Shape agent and/or stabilizer are avirulent for recipient under applied dosage and concentration, and it includes buffer, such as Phosphate, citrate and other organic acids；Antioxidant, including ascorbic acid, glutathione, cysteine, methionine And citric acid；Preservative (such as ethyl alcohol, benzyl alcohol, phenol, metacresol, parachlorometacresol, methyl p-hydroxybenzoate or to hydroxyl Yl benzoic acid propyl ester, benzalkonium chloride or combinations thereof object)；Amino acid, such as arginine, glycine, ornithine, lysine organize ammonia Acid, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenylalanine, tyrosine, tryptophan, methionine, silk Propylhomoserin, proline and combinations thereof；Monosaccharide, disaccharides and other carbohydrate；Low molecular weight (less than about 10 residues) polypeptide； Protein, such as gelatin or seralbumin；Chelating agent such as EDTA；Sugar, such as trehalose, sucrose, lactose, glucose, sweet dew Sugar, maltose, galactolipin, fructose, sorbose, gossypose, aminoglucose, N-METHYL-ALPHA-L-GLUCOSAMINE, galactosamine and neuraminic acid； And/or nonionic surfactant, such as tween, polyoxyethylene alkyl ether pluronic, Triton-X or polyethylene glycol (PEG)。
The pharmaceutical compositions can be liquid form, lyophilized form, or the liquid form reformulated by lyophilized form, In the lyophilized preparation reformulated before administration with sterile solution.Normal process for reformulating freeze-dried composition is add-back The pure water (volume removed during being generally equal to lyophilized) of certain volume；However, the solution comprising antibacterial agent can be used for producing use In the pharmaceutical compositions of parenteral administration；See also Chen (1992) Drug Dev Ind Pharm 18,1311-54.
Exemplary antibodies concentration in theme pharmaceutical compositions can range from about 1mg/mL to about 200mg/mL or about 50mg/mL to about 200mg/mL, or about 150mg/mL to about 200mg/mL.
The aqueous preparaton of the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure can be about in such as pH range 4.0 to about 7.0, or about 5.0 to about 6.0, or alternatively from about being prepared in 5.5 pH buffered solution.Suitable for this range The example of the buffer of interior pH includes phosphate-, histidine-, citrate-, succinate-, acetate-buffer and its His organic acid buffer liquid.The buffer concentration can be about 1mM to about 100mM, or about 5mM to about 50mM, this is depended on for example The buffer and the desired tension of the preparaton.
It may include tonicity agent in the antibody formulations to adjust the tension of the preparaton.Exemplary tonicity agent includes chlorination Sodium, potassium chloride, glycerol and any component from amino acid, sugar with and combinations thereof.In some embodiments, the water solubility Preparation be it is isotonic, although hypertonic or hypotonic solution can be it is suitable." isotonic " expression of term has to be compared with it Compared with some other solution equal tensions solution, such as saline or serum.It can be with about 5mM to the amount of about 350mM (such as with amount of 100mM to 350nM) uses tonicity agent.
Surfactant can also be added to the antibody formulations to reduce the aggregation of the antibody of the preparation and/or utmostly It reduces the formation of the particle in the preparaton and/or reduces absorption in ground.Exemplary surfactants include polyoxyethylene sorbitol Alcohol acid anhydride aliphatic ester (tween), polyoxyethylene alkyl ether (Brij), alkyl phenyl polyethylene oxides ether (Triton-X), polyoxy second Alkene-poiyoxypropylene copolymer (poloxamer, pluronic) and lauryl sodium sulfate (SDS).Suitable polyoxyethylene The example of sugar alcohol acid anhydride aliphatic ester is polysorbate20, (with trade mark polysorbas20TMSell) and polysorbate80 (spat with trade mark Temperature 80TMIt sells).The example of suitable Pluronic F68 is with titleF68 or pool Lip river are husky Nurse 188TMThe Pluronic F68 of sale.The example of suitable polyoxyethylene alkyl ether is with trade mark BRIJTM The polyoxyethylene alkyl ether of sale.The exemplary concentration of surfactant can range from about 0.001% to about 1%w/v.
Freeze drying protectant can be also added, to protect unstable active component (such as albumen) from unstable during freeze-drying The influence of condition.For example, as it is known that freeze drying protectant include sugared (including dextrose and saccharose)；Polyalcohol (including mannitol, mountain The pure and mild glycerol of pears)；With amino acid (including alanine, glycine and glutamic acid).It may include the freeze-drying of the amount of about 10mM to 500nM Protective agent.
In some cases, theme preparaton includes the anti-C1s antibody (the anti-C1s antibody of such as humanization) and one of the disclosure Kind or a variety of above confirmed medicaments (such as surfactant, buffer, stabilizer, tonicity agent), and essentially free of one Kind or Determination of Preservatives, such as ethyl alcohol, benzyl alcohol, phenol, metacresol, parachlorometacresol, methyl p-hydroxybenzoate or to hydroxyl Yl benzoic acid propyl ester, benzalkonium chloride or combinations thereof object.In other embodiments, preservative is with for example, about 0.001 to about 2% (w/v) concentration range is included in the preparaton.
For example, theme preparaton can be the liquid suitable for parenteral administration or the preparaton of freeze-drying, and may include: about The anti-C1s antibody (the anti-C1s antibody of such as humanization) of the disclosure of 1mg/mL to about 200mg/mL；About 0.001% to about 1% extremely A kind of few surfactant；The buffer of about 1mM to about 100mM；The stabilizer of optionally about 10mM to about 500mM；About 5mM To the tonicity agent of about 305mM；And the pH with about 4.0 to about 7.0.
Such as another example, the parenteral preparaton of theme is the preparation packet of the preparaton of liquid or freeze-drying, the liquid or freeze-drying Contain: the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of about 1mg/mL to about 200mg/mL；0.04% tween 20w/v；20mM L-Histidine；With 250mM sucrose；And with 5.5 pH.
Such as another example, the parenteral preparaton of theme includes the preparaton of freeze-drying, and the preparaton of the freeze-drying includes: 1) The theme antibody (such as anti-C1s antibody of humanization of the disclosure) of 15mg/mL；0.04% polysorbas20 w/v；20mM L-Histidine； With 250mM sucrose；And with 5.5 pH；Or 2) the theme antibody of 75mg/mL；0.04% polysorbas20 w/v；20mM L- group ammonia Acid；With 250mM sucrose；And with 5.5 pH；Or 3) the theme antibody of 75mg/mL；0.02% polysorbas20 w/v；20mM L- group Propylhomoserin；With 250mM sucrose；And with 5.5 pH；Or 4) the theme antibody of 75mg/mL；0.04% polysorbas20 w/v；20mM L- Histidine；With 250mM trehalose；And with 5.5 pH；Or 5) the theme antibody of 75mg/mL；0.02% polysorbas20 w/v；20mM L-Histidine；With 250mM trehalose；And with 5.5 pH.
Such as another example, the parenteral preparaton of theme is liquid adjustments, which includes: 1) 7.5mg/mL Theme antibody；0.02% polysorbas20 w/v；120mM L-Histidine；With 250 125mM sucrose；And with 5.5 pH；Or 2) The theme antibody of 37.5mg/mL；0.02% polysorbas20 w/v；10mM L-Histidine；With 125mM sucrose；And with 5.5 pH； Or 3) the theme antibody of 37.5mg/mL；0.01% polysorbas20 w/v；10mM L-Histidine；With 125mM sucrose；And with 5.5 pH；Or 4) the theme antibody of 37.5mg/mL；0.02% polysorbas20 w/v；10mM L-Histidine；125mM trehalose；And have 5.5 pH；Or 5) the theme antibody of 37.5mg/mL；0.01% polysorbas20 w/v；10mM L-Histidine；With 125mM trehalose； And with 5.5 pH；Or 6) the theme antibody of 5mg/mL；0.02% polysorbas20 w/v；20mM L-Histidine；With 250mM seaweed Sugar；With the pH with 5.5；Or 7) the theme antibody of 75mg/mL；0.02% polysorbas20 w/v；20mM L-Histidine；And 250mM Mannitol；And with 5.5 pH；Or 8) the theme antibody of 75mg/mL；0.02% polysorbas20 w/v；20mM L-His；With 140mM sodium chloride；With the pH with 5.5；Or 9) the theme antibody of 150mg/mL；0.02% polysorbas20 w/v；20mM L- group ammonia Acid；With 250mM trehalose；And with 5.5 pH；Or 10) the theme antibody of 150mg/mL；0.02% polysorbas20 w/v；20mM L-Histidine；With 250mM mannitol；With the pH with 5.5；Or 11) the theme antibody of 150mg/mL；0.02% polysorbas20 w/v； 20mM L-Histidine；With 140mM sodium chloride；And with 5.5 pH；Or 12) the theme antibody of 10mg/mL；0.01% tween 20w/v；20mM L-Histidine；With 40mM sodium chloride；With the pH with 5.5.
Theme antibody can be used in aerosol formulations to apply via sucking.Theme antibody can be made, which to be configured to pressurization, to be connect The propellant received, such as dicholorodifluoromethane, propane, nitrogen etc..Aerosol formulations (such as nasal spray formulation) include having Aqueous or other solution of the purified active agents of preservative and isotonic agent.It is extremely mutually simultaneous with schneiderian membrance to adjust such preparaton The pH of appearance and isotonic state.
It can provide unit dosage forms for oral administration, such as syrup, elixir and suspension, wherein each dosage unit The composition of (such as teaspoonful, the amount of a soupspoon or a piece of) containing predetermined amount.Similarly, for injection or vein The unit dosage forms of interior application can be in the composition comprising theme antibody as sterile water, physiological saline or another pharmaceutically acceptable The solution of carrier.
As used in this article, term " unit dosage forms " refers to the physics of the unit dose as suitable humans and animals subject Discrete unit is gone up, each unit for calculating the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure containing predetermined amount Amount, the amount are enough to generate and pharmaceutically acceptable diluent, carrier or the associated desired effect of medium.Theme antibody Specification may depend on drug effect associated with each antibody in applied specific antibodies and effect to be achieved and the host It learns.
Other methods of application will be also used in method of the invention.For example, theme antibody can be made to be configured to suppository, one In a little situations, it is configured to aerosol and intranasal compositions.For suppository, the vector composition will include conventional adhesive and Carrier, such as polyalkylene glycol or triglycerides.Such suppository can be by being about 0.5% to about 10% (w/w) containing range The mixture of the active component of (for example, about 1% to about 2%) is formed.
Intranasal preparation is typically included the medium for neither stimulating schneiderian membrance nor significantly destroying fibre function.It can apply Diluent, such as water, aqueous salt or other known substance.The intranasal formulations can also contain preservative, such as, but not limited to, Methaform and benzalkonium chloride.Surfactant may be present to enhance absorption of the schneiderian membrance to theme antibody.
Theme antibody can be used as injectable preparaton and be administered.In general, Injectable composition is made to be prepared into liquid solution Or suspension；The solid form of the solution being suitable in liquid vehicle or suspension before the injection can also be prepared.Said preparation is also It can be emulsified or be encapsulated in the antibody in liposome medium.
Suitable excipient vehicles are, for example, water, salt, dextrose, glycerol, ethyl alcohol etc. and its composition.In addition, such as Fruit needs, which can contain less amount of auxiliary substance, such as wetting agent or emulsifier or pH buffer.Prepare such dose The current methods of type are known to those skilled in the art, or will be obvious.See, e.g. Remington's Pharmaceutical Sciences,Mack Publishing Company,Easton, Pennsylvania, the 17th edition, 1985.Anyway, composition to be administered or preparaton, which will contain, is enough treating Patient in realize desired state amount theme antibody.
The pharmaceutical acceptable excipient (such as medium, adjuvant, carrier or diluent) is easy to public's acquisition.In addition, medicine Acceptable auxiliary substance, such as pH adjustment and buffer, tension adjustment agent, stabilizer, wetting agent etc. are learned, public's acquisition is easy to.
In some embodiments, the anti-C1s antibody (the anti-C1s antibody of such as humanization) of the disclosure is made to be configured to controlled release Preparation.Method known in the art can be used to prepare extended release preparation.The suitable example of extended release preparation includes containing Its mesostroma is the semipermeability base of the solid hydrophobic polymers of the antibody of the form (such as film or microcapsules) of molded article Matter.The example of sustained-release matrix includes polyester, the copolymer of Pidolidone and Pidolidone ethyl ester, nondegradable ethylene- Vinyl acetate, hydrogel, polyactide, degradable lactic acid-ethanol copolymer and poly- D- (-) -3-hydroxybutyrate.Pass through Using suitable additive, by control moisture content and by developing specific polymer matrix composition, can prevent include The bioactivity of antibody in extended release preparation can the loss of energy and immunogenicity possibility variation.
It is believed that the controlled release in the scope of the present disclosure refers to any one of many extended release dosage forms.For this Disclosed purpose, it is believed that following term is substantially equivalent to controlled release: continuous release (continuous release), by Controlled release is put (controlled release), sustained release (delayed release), drug storehouse storage (depot), delay release (extended release) gradually discharges (gradual release), releases immediately (immediate release), for a long time It discharges (long-term release), program-controlled release (programmed release), extended release (prolonged Release), (proportionate release) is discharged in proportion, delays release (protracted release), storage (repository), postpone (retard), slow release (slow release), interval release (spaced release) is held Continuous release (sustained release), time coat (time coat), time controlled released (timed release), delay are made With (delayed action), extension acts on (extended action), divides time effect (layered-time action), Long-acting (long acting), extension act on (prolonged action), and repeat function (repeated action) is slowly made With (slowing acting), continuous action (sustained action) and continuous action (sustained-action) medicine Object.Further discussing for these terms is found in LesczekKrowczynski,Extended-Release Dosage Forms, in 1987 (CRC Press LLCs).
Various controlled-release technologies cover very extensive pharmaceutical dosage form.Controlled-release technologies include but is not limited to department of physics System and chemical system.
Physical system includes but is not limited to the stocking system with rate controlling membranes, such as microencapsulation, big encapsulated and membrane system System；Stocking system without rate controlling membranes, such as doughnut, ultramicropore cellulose triacetate and porous polymer matrix Material and foam；Total system, including be physically dissolved in it is non-porous, polymerization or those of in elastomeric matrices system (for example, not Erodable, easily lose, environmental factor into leaching and it is degradable), and be physically dispersed in it is non-porous, polymerization or in elastomeric matrices Material (for example, not erodable, easily lose, environmental factor into leaching and degradable)；Laminar structure, including be chemically similar with external control layer Or dissimilar reservoir；With other physical methods, absorption in such as osmotic pumps or ion exchange resin.
Chemical system includes but is not limited to the chemical erosion (for example, heterogeneous or uniform erosion) of polymer substrate, or poly- The bioerosion (for example, heterogeneous or homogeneous) of polymer matrix.The additional discussion of the classification of system for controlled release can See Agis F.Kydonieus, Controlled Release Technologies:Methods, Theory and Applications,1980(CRC Press,Inc.)。
There are many controlled release drug preparatons that oral administration is used for through developing.These controlled release drug preparatons Including but not limited to control the stomach and intestine delivery system of osmotic pressure；Control the stomach and intestine delivery system of fluid pressure；Control the stomach of film infiltration Intestine delivery system comprising control the stomach and intestine delivery apparatus of microporous membrane permeation；The intestinal-specific controlled release stomach and intestine of anti-gastric juice are passed Send device；Control the stomach and intestine delivery system of gel diffusion；With control ion exchange stomach and intestine delivery system comprising cation and Anionic drugs.Additional information about controlled release drug delivery system is found in Yie W.Chien, Novel Drug Delivery Systems,1992(Marcel Dekker,Inc.)。
Dosage
Suitable dosage can be determined by attending physician or other qualified medical workers according to various clinical factors.Such as doctor Learn it is well known in the art, for any patient dosage depend on many factors, the figure including patient, body surface area, year Age, specific compound to be administered, the gender of patient, the time of application and approach, general health, and be administered simultaneously its His drug.The anti-C1s antibody (the anti-C1s antibody of such as humanization) of the disclosure can be with every dosage 1ng/kg weight and 20mg/kg body Amount application between weight, such as between 0.1mg/kg weight and 10mg/kg weight, such as 0.5mg/kg weight and 5mg/kg weight Between；However, the dosage below or above this exemplary range is to imagine, it is especially considering that above-mentioned factor.If scheme It is continuous infusion, then it can also be in the range of every 1 μ g to 10mg of kg body weight per minute.
In some embodiments, the dosage of the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure is in 0.001 μ In the range of g to 1000 μ g；However, the dosage below or above this illustrative range is to imagine, it is especially considering that above-mentioned Factor.In some embodiments, which can be for example, about 0.0001 to 100mg/kg, or about 0.01 to 5mg/kg (example Such as 0.02mg/kg, 0.25mg/kg, 0.5mg/kg, 0.75mg/kg, 1mg/kg, 2mg/kg etc.) range of weight.For example, dosage It can be 1mg/kg weight or 10mg/kg weight or in the range of 1-10mg/kg, or at least 1mg/kg.In above range Dosage median is also intended in the scope of the present disclosure.
In some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure is to provide about 1 μ g/ml Amount to the peak serum concentration of about 1mg/ml is administered, for example, about 1 μ g/ml to about 2.5 μ g/ml, about 2.5 μ g/ml to about 5 μ g/ml, about 5 μ g/ml are to about 7.5 μ g/ml, about 7.5 μ g/ml to about 10 μ g/ml, about 10 μ g/ml to about 25 μ g/ml, about 25 μ g/ Ml to about 50 μ g/ml, about 50 μ g/ml are to about 100 μ g/ml, about 100 μ g/ml to about 250 μ g/ml, about 250 μ g/ml to about 500 μ G/ml, about 500 μ g/ml are to about 750 μ g/ml, about 750 μ g/ml to about 1000 μ g/ml.In some embodiments, theme is anti- C1s antibody is administered with the amount for providing the peak serum concentration for being greater than 1mg/ml, for example, about 1mg/ml to about 2mg/ml, about 2mg/ml to about 5mg/ml, about 5mg/ml are to about 10mg/ml.The humanized antibody of the disclosure can be applied according to any planning chart With, or can apply up to any time section.
The person skilled in the art will easily understand dosage level and application planning chart can be according to the function of antibody specific, diseases The seriousness of shape and the subject change the sensibility of side effect.For the preferred dosage of appointed compound and application Planning chart can be readily determined by various modes by those skilled in the art.
The approach of application
Using any available method and approach for being suitable for drug delivery, including internal and in vitro method and whole body With the application of topic route, theme antibody is applied to individual.
Conventional and pharmaceutically acceptable administration method includes intranasally that intramuscular is intratracheally, intrathecal, encephalic, subcutaneously, skin It is interior, it is local, intravenously, and in peritonaeum, intra-arterial (such as via arteria carotis), spinal cord or brain delivery, rectum, intranasal, it takes orally And other enteral and parenteral administration method.Administration method can be combined, and if necessary, or be adjusted, this Depending on the antibody and/or desired effect.Theme antibody compositions can be applied with single dose or multiple dosage.In some realities It applies in scheme, theme antibody compositions are administered orally.In some embodiments, theme antibody compositions are applied via inhalation route With.In some embodiments, theme antibody compositions intranasal administration.In some embodiments, theme antibody compositions office Portion's application.In some embodiments, theme antibody compositions encephalic is applied.In some embodiments, theme antibody combination Object is intravenously applied.In some embodiments, theme antibody compositions subcutaneous administration.In some embodiments, theme is anti- The application of body composition intramuscular.In some embodiments, the intrathecal application of theme antibody compositions.
Using any available conventional method and approach for being suitable for Conventional drug delivery, the approach including whole body or part, The antibody of the disclosure is applied to host.In general, the administration method as expected from the disclosure includes but is not limited to enteral, stomach and intestine Outside or inhalation route.
Parenteral administration method is including but not necessarily limited to local other than sucking application, transdermal, subcutaneously, intramuscular, eye It is intracapsular in socket of the eye, it is intraspinal, in breastbone, intrathecal and intravenous route, i.e., except through alimentary canal outside any administration method.It can Parenteral administration is carried out to realize the whole body or local delivery of theme antibody.In the case where needing systemic delivery, application is usual It is related to invasive or systemic Absorption part or the mucosal administration of pharmaceutical preparation.
It can also be by enteral administration by theme antibody delivery to the subject.Enteral administration path includes but is not limited to mouth Clothes and rectum (such as using suppository) delivering.
Being referred to by treatment at least improves symptom relevant to the pathology patient's condition of host pain is made, wherein improving in broad sense It is upper for refer at least reduce relevant to the pathology patient's condition (such as disease or illness of complement-mediated) for being treated parameter (such as Symptom) magnitude.Therefore, treatment further includes following situations, wherein the pathology patient's condition, or at least relative symptom, has been Inhibit entirely, such as prevents to occur, or stopped, such as terminated, so that the host no longer suffers from the pathology patient's condition, or Symptom at least characterized by the pathology patient's condition.
In some embodiments, by injecting the anti-C1s antibody of the disclosure (such as the anti-C1s of humanization and/or delivering Antibody) it applies the site (for example) into cerebral artery or is applied directly in brain tissue.It can also be by theme antibody (such as humanization Antibody) it is applied directly to target site, such as the target site is delivered to by biolistic (biolistic).
According to the subject methods, various hosts (wherein term " host " and term " subject ", " individual ", and " patient " Use interchangeably herein) it is medicable.In general, such host is " mammal (mammal) " or " mammal (mammalian) ", wherein these terms are widely used in the organism within description mammal, including carnivore mesh (example Such as cat), herbivore mesh (such as ox, horse and sheep) is eaten miscellaneous animal mesh (such as dog, goat and pig), Rodentia (such as it is small Mouse, cavy and rat) and primate (such as people, chimpanzee and monkey).In some embodiments, which is to have to mend The individual of system system, such as mammal, fish or invertebrate.In some embodiments, which is containing complement system The mammal of system, fish or invertebrate pet, agricultural animal, labour use animal, zoo animal or experimental animal.One In a little embodiments, which is people.
The embodiment includes comprising being suitable for containing the composition comprising the anti-C1s antibody of theme for being applied to individual The composition of container.It is suitable in the container containing pharmaceutical compositions for example, theme antibody can be placed in.The container can be, example Such as, bottle (such as with locking device, such as lid), bubble device (such as it can provide each bubble-cap one or more dosage Encapsulating), phial, the soft package mylar or polybag of sealing (for example), ampoule is (for the single dose in solution Amount), dropper, syringe, film, pipe etc..In some embodiments, container (such as sterile chamber) includes theme pharmaceutical composition Object.In some embodiments, which is bottle or syringe.In some embodiments, which is bottle.In some implementations In scheme, which is syringe.
Anti- C1s antibody (such as the people for the disclosure for having unit dose (for example, with oral or injectable dosage) is provided The anti-C1s antibody of sourceization) kit.In such kit, in addition to the container containing the unit dose, it will also contain and be described The antibody is in the information-package inset for treating purposes and adjoint benefit in the interested pathology patient's condition.Preferred compound and Unit dose is those described above compound and unit dose.
Treat the disease of complement-mediated or the method for illness
The vaccination that the disclosure provides the anti-C1s antibody disclosed in anti-C1s antibody or code book is in need tested The method of person.In some embodiments, this method includes treating the disease or illness of complement-mediated.This method, which is usually directed to, to be applied With the anti-C1s antibody (such as the anti-C1s antibody of humanization) of a effective amount of disclosure, or the pharmaceutical compositions comprising such antibody are extremely Individual in need.In some cases, the anti-C1s antibody of application theme adjusts the cell of individual, organizes, in liquid or organ Complement C1s activity, and treat the disease or illness of the complement-mediated.
The some aspects of the disclosure provide the method for inhibiting the activation of complement component C4 in individual, and this method includes that application has The anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of effect amount, or the pharmaceutical compositions comprising such antibody to this Body.The disclosure, which provides, inhibits the active method of complement C1s in individual, and this method includes applying a effective amount of disclosure to the individual Anti- C1s antibody (such as the anti-C1s antibody of humanization) or the pharmaceutical compositions comprising such antibody.The disclosure, which provides, reduces individual In (such as the liquid in individual, tissue or organ in) complement component cleaved products horizontal method, this method includes pair The anti-C1s antibody (such as the anti-C1s antibody of humanization) or the pharmacy comprising such antibody that the individual applies a effective amount of disclosure Composition.
In some cases, it includes to this that the treatment of the disclosure, which has the disease of complement-mediated or the method for the individual of illness, Individual applies the anti-C1s antibody (such as the anti-C1s antibody of humanization) or a effective amount of pharmaceutical compositions of a effective amount of disclosure, should Pharmaceutical compositions include: a) the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure；And b) it is suitable for application to such The pharmaceutical acceptable excipient of body.In some embodiments, which is mammal.In some embodiments, this Body is people.Application can pass through any approach well known by persons skilled in the art, including approach disclosed herein.In some implementations In scheme, application is intravenous.In some embodiments, application is intrathecal.In some embodiments, application is skin Under.In some embodiments, application is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The horizontal amount of complement component cleaved products.In some cases, anti-C1s antibody (such as the source of people of the disclosure of " effective quantity " Change anti-C1s antibody), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmacy Composition is when it is applied to individual in need with one or more dosage, make in the individual (such as in the individual Liquid, in tissue or organ) levels of complement component cleaved products reduces at least about 1%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% amount, phase Compared with the liquid in the case where lacking with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), tissue or device The level of complement component cleaved products in official.In some embodiments, which is mammal.In some embodiments In, which is people.Application can pass through any approach well known by persons skilled in the art, including approach disclosed herein.? In some embodiments, application is intravenous.In some embodiments, which is intrathecal.In some implementations In scheme, which is intravenous.In some embodiments, which is subcutaneous.In some embodiment party In case, which is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The active amount of classic complement approach.In some cases, (such as humanization is anti-for the anti-C1s antibody of the disclosure of " effective quantity " C1s antibody), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical composition Object is when it is applied to individual in need with one or more dosage, in about 48 hours for applying the anti-C1s antibody, about In 24 hours, in about 12 hours, in about 8 hours, or in about 4 hours, make in the individual (such as the liquid in the individual, group Knit or organ in) the activity of classic complement approach reduce at least about 1%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% amount, compared to lacking use Liquid in the case where the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), classical in tissue or organ are mended The activity of body approach.In some embodiments, which is mammal.In some embodiments, which is people.It applies Any approach well known by persons skilled in the art, including sheet can be passed through with the anti-C1s antibody (such as the anti-C1s antibody of the humanization) Approach disclosed herein.In some embodiments, which is intrathecal.In some embodiments, application way Diameter is intravenous.In some embodiments, which is subcutaneous.In some embodiments, the administration method It is intramuscular.Any one of various methods can be used to be measured for the activity level of classic complement approach.It is non-as one Limitative examples, the activity of classic complement approach can isolated measuring, for example, be obtained from the blood of the individual by measurement, serum, or The activity level of classic complement approach in plasma sample.For example, the classic complement approach in blood, serum or plasma sample Can activated ex vivo, and the amount of the complement component cleaved products (such as C5b-9) generated by this Class Activation can be measured.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The active amount of classic complement approach.In some cases, (such as humanization is anti-for the anti-C1s antibody of the disclosure of " effective quantity " C1s antibody), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical composition Object is when it is applied to individual in need with one or more dosage, in about 48 hours for applying the anti-C1s antibody, about In 24 hours, in about 12 hours, in about 8 hours, or in about 4 hours, make in the individual (such as the liquid in the individual, group Knit or organ in) the activity level of classic complement approach reduce at least about 1%, at least about 5%, at least about 10%, at least About 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least About 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% amount, compared to lacking Liquid in the case where with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), tissue or organ in classics The activity level of complement pathway.In some embodiments, which is mammal.In some embodiments, the individual It is people.Applying the anti-C1s antibody (such as the anti-C1s antibody of the humanization) can be by any way well known by persons skilled in the art Diameter, including approach disclosed herein.In some embodiments, which is intrathecal.In some embodiments, The administration method is intravenous.In some embodiments, administration method is subcutaneous.In some embodiments, this is applied It is intramuscular with approach.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The active amount of classic complement approach.In some cases, (such as humanization is anti-for the anti-C1s antibody of the disclosure of " effective quantity " C1s antibody), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical composition Object is when it is applied to individual in need with one or more dosage, maintain in the individual (such as the liquid in the individual Body, in tissue or organ) activity level at least about 1% of classic complement approach, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% reduced amount, compared to Liquid in the case where lacking with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), tissue or organ in The activity level of classic complement approach, wherein maintaining the reduction about 4 hours to about 30 days, (such as 4 hours to 8 hours, 8 hours extremely 24 hours, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days, 14 days to 21 days or 21 days to 30 days) period.In some realities It applies in scheme, which is mammal.In some embodiments, which is people.Applying the anti-C1s antibody (such as should The anti-C1s antibody of humanization) any approach well known by persons skilled in the art, including approach disclosed herein can be passed through.One In a little embodiments, which is intrathecal.In some embodiments, which is intravenous.Some In embodiment, which is subcutaneous.In some embodiments, which is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The active amount of classic complement approach.In some cases, (such as humanization is anti-for the anti-C1s antibody of the disclosure of " effective quantity " C1s antibody), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical composition Object is when it is applied to individual in need with one or more dosage, maintain in the individual (such as the liquid in the individual Body, in tissue or organ) activity level at least about 1% of classic complement approach, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% reduced amount, compared to Liquid in the case where lacking with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), tissue or organ in The activity level of classic complement approach, wherein maintaining the reduction about 4 hours to about 21 days, (such as 4 hours to 8 hours, 8 hours extremely 24 hours, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days or 14 days to 21 days) period.In some embodiments, should Individual is mammal.In some embodiments, which is people.Apply the anti-C1s antibody (such as the anti-C1s of the humanization Antibody) any approach well known by persons skilled in the art, including approach disclosed herein can be passed through.In some embodiments In, which is intrathecal.In some embodiments, which is intravenous.In some embodiments, The administration method is subcutaneous.In some embodiments, which is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The horizontal amount of middle complement component cleaved products.In some cases, anti-C1s antibody (such as the people of the disclosure of " effective quantity " The anti-C1s antibody of sourceization), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme medicine Learning composition is to apply the anti-C1s antibody (such as the people when it is applied to individual in need with one or more dosage The anti-C1s antibody of sourceization) about 48 hours in, in about 24 hours, in about 12 hours, in about 8 hours, or in about 4 hours, make this The horizontal of complement component cleaved products reduces at least about in (such as the liquid in the individual, tissue or organ in) in body 1%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% amount, compared in the case where lacking with the anti-C1s Antybody therapy (such as with the anti-C1s antibody Before treatment) liquid, tissue or the complement component cleaved products in organ level.In some embodiments, which is Mammal.In some embodiments, which is people.Applying the anti-C1s antibody (such as the anti-C1s antibody of the humanization) can Pass through any approach well known by persons skilled in the art, including approach disclosed herein.In some embodiments, the application Approach is intrathecal.In some embodiments, which is intravenous.In some embodiments, application way Diameter is subcutaneous.In some embodiments, which is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The horizontal amount of middle complement component cleaved products.In some cases, anti-C1s antibody (such as the people of the disclosure of " effective quantity " The anti-C1s antibody of sourceization), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme medicine Learning composition is to apply the about 48 small of the anti-C1s antibody when it is applied to individual in need with one or more dosage When it is interior, in about 24 hours, in about 12 hours, in about 8 hours, or in about 4 hours, make in the individual (such as in the individual Liquid, in tissue or organ) the horizontal of complement component cleaved products reduce at least about 1%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% amount, phase Compared with liquid in the case where lacking with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), tissue or device The level of complement component cleaved products in official.In some embodiments, which is people.Apply the anti-C1s antibody (such as The anti-C1s antibody of the humanization) any approach well known by persons skilled in the art, including approach disclosed herein can be passed through.? In some embodiments, which is intrathecal.In some embodiments, which is intravenous.One In a little embodiments, which is subcutaneous.In some embodiments, which is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The horizontal amount of middle complement component cleaved products.In some cases, anti-C1s antibody (such as the people of the disclosure of " effective quantity " The anti-C1s antibody of sourceization), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme medicine Learning composition is when it is applied to individual in need with one or more dosage, maintain in the individual (such as in the individual In liquid, in tissue or organ) the horizontal of complement component cleaved products reduce at least about 1%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% amount, phase Compared with liquid in the case where lacking with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), tissue or The level of the complement component cleaved products in organ, wherein maintaining the reduction about 4 hours to about 30 days (such as 4 hours to 8 small When, 8 hours to 24 hours, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days, 14 days to 21 days, 21 days to 30 days) period. In some embodiments, which is mammal.In some embodiments, which is people.Apply the anti-C1s antibody (such as the anti-C1s antibody of the humanization) can pass through any approach well known by persons skilled in the art, including way disclosed herein Diameter.In some embodiments, which is intrathecal.In some embodiments, which is intravenous. In some embodiments, which is subcutaneous.In some embodiments, which is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or It when multiple dosage are applied to individual in need, reduces in the individual (such as the liquid in the individual, tissue or organ in) The horizontal amount of middle complement component cleaved products.In some cases, anti-C1s antibody (such as the people of the disclosure of " effective quantity " The anti-C1s antibody of sourceization), or " effective quantity " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme medicine Learning composition is when it is applied to individual in need with one or more dosage, maintain in the individual (such as in the individual In liquid, in tissue or organ) levels at least about 1% of complement component cleaved products, at least about 5%, at least about 10%, At least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, At least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% reduced amount, phase Compared with liquid in the case where lacking with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy), tissue or device The level of the complement component cleaved products in official, wherein maintaining the reduction about 4 hours to about 21 days (such as 4 hours to 8 small When, 8 hours to 24 hours, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days or 14 days to 21 days) period.In some implementations In scheme, which is mammal.In some embodiments, which is people.Apply the anti-C1s antibody (such as the people The anti-C1s antibody of sourceization) any approach well known by persons skilled in the art, including approach disclosed herein can be passed through.Some In embodiment, which is intrathecal.In some embodiments, which is intravenous.In some realities It applies in scheme, which is subcutaneous.In some embodiments, which is intramuscular.
In some cases, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure of " effective quantity ", or " effectively Amount " the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure theme pharmaceutical compositions be work as its with one or When multiple dosage are applied to individual in need, make in the individual (such as the liquid in the individual, tissue or organ in) C4b2a (i.e. Complement C4 b and C2a compound；Also referred to as " C3 convertase ") generation reduce at least about 1%, at least about 5%, until Few about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, until Few about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% Amount, compared in the case where lacking with the anti-C1s Antybody therapy (such as before with the anti-C1s Antybody therapy) individual (or The liquid, tissue or organ) in generate C4b2a amount.In some embodiments, which is mammal.Some In embodiment, which is people.Application can be including disclosed herein by any approach well known by persons skilled in the art Approach.In some embodiments, application is intravenous.In some embodiments, which is intrathecal.One In a little embodiments, which is intravenous.In some embodiments, which is subcutaneous.Some In embodiment, which is intramuscular.
The disclosure provides the method for adjusting complement activation.In some embodiments, this method inhibits complement activation, such as To reduce the generation of C4b2a.In some embodiments, the disclosure provides for adjusting the disease or illness with complement-mediated The method of complement activation in body, this method include anti-C1s antibody (such as the anti-C1s of humanization to the individual application disclosure Antibody) or the disclosure pharmaceutical compositions, wherein the pharmaceutical compositions include that (such as humanization is anti-for the anti-C1s antibody of the disclosure C1s antibody).In some embodiments, such method inhibits complement activation.In some embodiments, which is lactation Animal.In some embodiments, which is people.Application can by any approach well known by persons skilled in the art, including Approach disclosed herein.In some embodiments, application is intravenous.In some embodiments, application is intrathecal 's.In some embodiments, application is subcutaneous.In some embodiments, which is intramuscular.
The disease or illness of complement-mediated are the cells that its characterization is individual, are organized, abnormal amount in liquid or organ The illness of complement C1s or the complement C1s proteolytic activity of abnormal level.
In some cases, the disease of complement-mediated or the characterization of illness are to be present in cell, in tissue or liquid The complement C1s activity for improving and (being higher than normal) C1s of amount or improve the standard.For example, in some cases, the disease of complement-mediated Or the characterization of illness be the raising amount being present in brain tissue and/or cerebrospinal fluid and/or improve active C1s.Cell, group Knit or liquid in the C1s of " be higher than normal " amount indicate the cell, the amount of C1s is higher than normal, control level in tissue or liquid, Such as normal, the control level higher than individual or group in same age group individual.Cell is organized, in organ or fluid " higher than normal " horizontal C1s activity instruction by the cell, is organized, and the proteolysis of the C1s effect in organ or fluid is cut It cuts and is higher than normal, control level, such as higher than normal, the control level of individual or group in same age group individual.One In a little situations, the individual of disease or illness with complement-mediated shows one or more additional diseases of such disease or illness Shape.
In other situations, the disease of complement-mediated or the characterization of illness are to be present in cell, in tissue or liquid Lower than the C1s of normal amount or the complement C1s activity of reduced levels.For example, in some cases, the disease or illness of complement-mediated Characterization be the relatively low amount being present in brain tissue and/or cerebrospinal fluid and/or lower active C1s.Cell, tissue or liquid " lower than normal ", the C1s of amount indicates the cell in body, and the amount of C1s is lower than normal, control level in tissue or liquid, such as less than Normal, the control level of individual or group in same age group individual.Cell, " lower than normal " is horizontal in tissue or liquid The instruction of C1s activity by the cell, the proteolysis cutting of the C1s effect in tissue or liquid lower than normal, control level, Normal, the control level of individual or group such as less than in same age group individual.In some cases, there is complement-mediated Disease or the individual of illness show one or more additional symptoms of such disease or illness.
The disease or illness of complement-mediated are the diseases that wherein amount of complement C1s or activity lead to disease or illness in individual Disease or illness.In some embodiments, the disease of the complement-mediated or illness are selected from the group being made up of: isoimmunization disease Disease, autoimmune disease, cancer, hematologic disease, communicable disease, diseases associated with inflammation, ischemical reperfusion injury, neurological Property disease, Neurodegenerative conditions, eye disease, kidney trouble, graft rejection, vascular diseases and vasculitis diseases.One In a little embodiments, the disease or illness of the complement-mediated are autoimmune diseases.In some embodiments, which is situated between The disease or illness led are isoimmunization diseases.In some embodiments, the disease of the complement-mediated or illness are cancers.? In some embodiments, the disease or illness of the complement-mediated are communicable diseases.In some embodiments, the complement-mediated Disease or illness be diseases associated with inflammation.In some embodiments, the disease of the complement-mediated or illness are hematologic diseases.? In some embodiments, the disease or illness of the complement-mediated are ischemical reperfusion injuries.In some embodiments, the complement The disease or illness of mediation are eye diseases.In some embodiments, the disease of the complement-mediated or illness are kidney troubles. In some embodiments, the disease of the complement-mediated or illness are graft rejection.In some embodiments, which is situated between The disease or illness led are antibody-mediated graft rejection.In some embodiments, the disease or illness of the complement-mediated It is vascular diseases.In some embodiments, the disease of the complement-mediated or illness are vasculitis illnesss.In some embodiments In, the disease or illness of the complement-mediated are neurodegenerative disease or illness.In some embodiments, the complement-mediated Disease is neurodegenerative disease.In some embodiments, the illness of the complement-mediated is Neurodegenerative conditions.
The disease of complement-mediated or the example of illness include but is not limited to age related macular degeneration, Alzheimers Disease, amyotrophic lateral sclerosis, allergic reaction, argyrophilic grain dementia, arthritis (such as rheumatoid arthritis), asthma, Atherosclerosis, atypia hemolytic uremic syndrome, autoimmune disease (including for example Autoimmune hemolytic is poor Blood (AIHA)；Tepid-type AIHA；Mixed type AIHA；Deng), this syndrome of Braak that-simon, Behcet's disease, Britain's type shallow lake Powder Angiopathy, bullous pemphigoid, Buerger's disease, C1q nephrosis, cancer, catastrophic antiphospholipid syndrome, brain amyloid blood Pipe disease, cold coagulation disease, corticobasal degeneration, Creutzfeldt-Jakob disease, Crohn's disease, cryoglobulinemia Property vasculitis, boxer's dementia, dementia with Lewy body (DLB), with calcification Diffuse neurofibrillary tangle, plate-like erythema wolf Sore, Down syndrome, angstrom Wen's syndrome, focal segmental glomerulosclerosis, formal thought disorder, Frontotemporal dementia (FTD), with the chain Frontotemporal dementia with Parkinson's disease of chromosome 17, frontotemporal lobar degeneration, Gerstmann- Straussler-Scheinker disease, actue infectious polyradiculoneuritis, Hallervorden Spatz disease, hemolytic uremic syndrome, Hereditary angioedema, hypophosphatemia, idiopathic pneumonia syndrome, immune complex disease, inclusion body myositis, infectiousness Disease (for example, by bacterium (such as Neisseria meningitidis or streptococcus)), viral (such as human immunodeficiency virus (HIV)), Or disease caused by other infectious agents), diseases associated with inflammation, ischemia/reperfusion injury, mild cognitive impairment, immunity blood is small Plate reduction property purpura (ITP), A type deficiency of molybdenum cofactor disease (MoCD), I type membrano proliferative glomerulonephritis (MPGN), II type film Hyperplastic glomerular nephritis (MPGN) (compactness storage disorders), membraneous nephritis, multiple infarct dementia, lupus (such as systematicness Lupus erythematosus (SLE)), glomerulonephritis, Kawasaki disease, multifocal motor neuropathy, multiple sclerosis, multi-system atrophy, severe Myasthenia, myocardial infarction, myotonia dystrophy, neuromyelitis optica, c-type Niemann-Pick disease are twined with nerve fibril Non- Guam type motor neuron disease of knot, Parkinson's disease, with dull-witted Parkinson's disease, paroxysmal nocturnal hemoglobin Urine, pemphigus vulgaris, Pick's disease, postencephalitic parkinsonism, polymyositis, prion protein cerebral amyloid angiopathy, into Gliosis under row cortex, stein-leventhal syndrome, psoriasis, septicemia, shiga toxin Escherichia coli (STEC)- HuS, Duchenne-Arandisease, apoplexy, subacute sclerosing panencephalitis, only entanglement type is dull-witted, graft rejection, vasculitis (such as ANCA relevant blood vessel is scorching), Wei Genashi granulomatosis, drepanocytosis, cryoglobulinemia, mixed type cryoglobulinemia, Idiopathic mixed type cryoglobulinemia, II type mixed type cryoglobulinemia, type III mixed type cryoglobulinemia, ephritis, Drug-induced thrombopenia, lupus nephritis, acquired epidermolysis bullosa, delaying type hemolytic blood transfusion reaction, Hypocomplementemia nettle rash vasculitic syndrome, false bullous keratopathy, blood platelet refractoriness, chronic inflammation demyelinate Property polyneuropathy (CIDP), myelodysplastic syndrome (MDS), Miller fisher's syndrome, acute inflammatory demyelinating Polyneuropathy (AIDP), acute exercise axon nerve is sick (AMAN), and acute exercise and feeling axon nerve are sick (AMSAN), With pharynx-neck-arm upper arm variant.In one embodiment, the disease of the complement-mediated or illness include bullous pemphigoid.? In one embodiment, the disease or illness of the complement-mediated include cold coagulation disease.In one embodiment, which is situated between The disease or illness led include autoimmune hemolytic anemia (AIHA).In one embodiment, the disease of the complement-mediated Disease or illness include immunologic thrombocytopenic purpura (ITP).In one embodiment, the disease or disease of the complement-mediated Disease includes multifocal motor neuropathy.In one embodiment, the disease of the complement-mediated or illness include optic nerve spinal cord It is scorching.
In some embodiments, the disease of the complement-mediated or illness include Alzheimer's disease.In some implementations In scheme, the disease or illness of the complement-mediated include Parkinson's disease.In some embodiments, the disease of the complement-mediated Or illness includes graft rejection.In some embodiments, the disease of the complement-mediated or illness are antibody-mediated transplanting Object repels.
In some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure prevents or delays individual The breaking-out of at least one symptom of the disease or illness of middle complement-mediated.In some embodiments, the anti-C1s antibody of the disclosure Reduce or eliminate at least one symptom of the disease of complement-mediated or illness in individual.The example of symptom include but is not limited to and from Body immunity disease, cancer, hematologic disease, communicable disease, diseases associated with inflammation, ischemic damage and reperfusion damage, nervus retrogression Disease, Neurodegenerative conditions, kidney trouble, graft rejection, eye disease, vascular diseases or vasculitis illness are relevant Symptom.The symptom can be nervous symptoms, for example, impaired cognitive function, memory disorders, motor function loss etc..The symptom It can also be the cell of individual, the activity of C1s albumen in tissue or liquid.The symptom can also be the cell of individual, organize, Or in liquid complement activation degree.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) to the individual for applying the disclosure is adjusted Individual cell, tissue or liquid in complement activation.In some embodiments, the application anti-C1s antibody of theme to individual inhibits Individual cell, tissue or liquid in complement activation.For example, in some embodiments, when it is made with one or more dosage When being applied to the individual of disease or illness with complement-mediated for monotherapy or with combination treatment, the anti-C1s antibody (example of theme Such as the anti-C1s antibody of humanization) inhibit complement activation at least about 1% in the individual, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95% or 100%, compared to before the anti-C1s Antybody therapy Complement activation in body.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) the reduction C3 of the disclosure is deposited to red On cell；For example, in some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure reduces C3b, The deposition on RBC such as iC3b).In some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure Inhibit the erythrocyte splitting of complement-mediated.
In some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure reduces C3 and deposits to blood On platelet；For example, in some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure reduces C3b, The deposition on blood platelet such as iC3b).
In some embodiments, apply the disclosure anti-C1s antibody (such as the anti-C1s antibody of humanization) cause selected from by The result of group consisting of: complement activation (a) is reduced；(b) improve cognitive function；(c) neurone loss is reduced；(d) it reduces Microglia activation；(e) lymphocytic infiltration is reduced；(f) macrophages infiltration is reduced；(g) antibody deposition is reduced, (h) is reduced Spongiocyte loss；(i) oligodendrocyte loss is reduced；(j) Dendritic Cells Infiltration is reduced；(k) neutrophil infiltration is reduced； (l) erythrocyte splitting is reduced；(m) red blood cell phagocytosis is reduced；(n) blood platelet phagocytosis is reduced；(o) blood platelet is reduced to split Solution；(p) improve transplanting object survival rate；(q) macrophage-mediated phagocytosis is reduced；(r) improve eyesight；(s) improve fortune Dynamic control；(t) improve thrombosis；(u) improve blood coagulation；(v) improve renal function；(w) antibody-mediated complement activation is reduced； (x) complement activation that autoantibody mediates is reduced；(y) improve anaemia；(aa) demyelinate is reduced；(ab) eosinophil is reduced Increase disease；(ac) it reduces C3 and deposits on red blood cell (such as reducing C3b, the deposition on RBC such as iC3b)；(ad) reduces C3 (such as reducing C3b, the deposition on blood platelet such as iC3b) is deposited on blood platelet；(ae) reduces anaphylatoxin toxin and produces It is raw；(af) blister that autoantibody mediates is reduced to be formed；(ag) itch of autoantibody induction is reduced；(ah) autoantibody is reduced The lupus erythematosus of induction；(ai) skin erosion that autoantibody mediates is reduced；(aj) it reduces since the red blood cell of transfusion reaction is broken It is bad；(ak) erythrocyte splitting due to allo-antibody is reduced；(al) haemolysis due to transfusion reaction is reduced；(am) it is anti-to reduce itself The platelet lysates that body mediates；(an) platelet lysates due to transfusion reaction are reduced；(ao) mast cells activation is reduced；(ap) Reduce mast cell histamine release；(aq) vasopermeability is reduced；(ar) oedema is reduced；(as) complement deposit is reduced in graft On endothelium；(at) anaphylatoxin in graft endothelium is reduced to generate；(au) separation of dermal-epidermal junction is reduced；(av) it reduces The generation of dermal-epidermal junction anaphylatoxin；(aw) complement activation that autoantibody mediates in graft endothelium is reduced；(ax) Reduce the loss of antibody-mediated neuromuscular junction；(ay) complement activation of neuromuscular junction is reduced；(az) mind is reduced Generation through the anaphylatoxin at neuromuscular junction；(ba) complement deposit of neuromuscular junction is reduced；(bb) paralysis is reduced； (bc) it reduces numb；(bd) the bladder control improved；(be) the intestines control improved；(bf) death relevant to autoantibody is reduced Rate；(bg) disease incidence relevant to autoantibody is reduced；(bh) schwann cell damage is reduced；(bi) schwann cell loss is reduced； (bj) damage of motoneurons is reduced；(bk) loss of motor neuron axon is reduced；(bl) improve action potential block； (bm) improve upper limb or lower extremity movement；(bn) improve neuron sensorimotor defect, and (bo) any combination thereof.
In some embodiments, tool is applied to when it using one or more dosage as monotherapy or with combination treatment Have complement-mediated disease or illness individual when, anti-C1s antibody (such as the anti-C1s antibody of theme) is effectively realized one or more At least about the 1% of following results, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, until Few about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or Reduction more than 90%: (a) complement activation；(b) decline of cognitive function；(c) neurone loss；(d) microglia activation； (e) lymphocytic infiltration；(f) macrophages infiltration；(g) antibody deposition, (h) spongiocyte loses；(i) oligodendrocyte loses； (j) Dendritic Cells Infiltration；(k) neutrophil infiltration；(l) erythrocyte splitting；(m) red blood cell phagocytosis；(n) blood platelet Phagocytosis；(o) platelet lysates；(p) graft rejection；(q) macrophage-mediated phagocytosis；(r) vision loss； (s) antibody-mediated complement activation；(t) complement activation that autoantibody mediates；(u) demyelinate；(v) eosinophilia Disease；(w) or any combination thereof, compared in the level or degree with the result in the individual before the anti-C1s Antybody therapy.
In some embodiments, tool is applied to when it using one or more dosage as monotherapy or with combination treatment Have complement-mediated disease or illness individual when, the anti-C1s antibody of theme effectively realizes one or more following results extremely Few about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least About 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or changing more than 90% It is kind: a) cognitive function；B) Graft survival rate；C) eyesight；D) motion control；E) thrombosis；F) blood coagulation；G) renal function；h) Hematocrit (red blood cell count(RBC))；And i) any combination thereof, compared to the knot in the individual before the anti-C1s Antybody therapy The level or degree of fruit.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) to the individual for applying the disclosure reduces Complement activation in the individual.For example, in some embodiments, when it is using one or more dosage as monotherapy or with group When conjunction therapy is applied to the individual of disease or illness with complement-mediated, the anti-C1s antibody of theme reduces complement in the individual and swashs Living at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, At least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, Compared to the complement activation in the individual before the anti-C1s Antybody therapy.
In some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) for applying the disclosure improves the individual In cognitive function.For example, in some embodiments, when it is treated using one or more dosage as monotherapy or with combining When method is applied to the individual of disease or illness with complement-mediated, the anti-C1s antibody of theme improves in the individual cognitive function extremely Few about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least About 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, compare The cognitive function in individual before the Yu Yong anti-C1s Antybody therapy.
In some embodiments, the anti-C1s antibody (such as the anti-C1s antibody of humanization) for applying the disclosure reduces the individual The rate of descent of middle cognitive function.For example, in some embodiments, when its using one or more dosage as monotherapy or with When combination treatment is applied to the individual of disease or illness with complement-mediated, the anti-C1s antibody of theme reduces to be recognized in the individual The rate of descent of function at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, At least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, Or more than 90%, compared in the rate of descent with the cognitive function in the individual before the anti-C1s Antybody therapy.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) to the individual for applying the disclosure reduces Neurone loss in the individual.For example, in some embodiments, when its using one or more dosage as monotherapy or with When combination treatment is applied to the individual of disease or illness with complement-mediated, the anti-C1s antibody of theme reduces neural in the individual Member loss at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or be more than 90%, compared to the neurone loss in the individual before the anti-C1s Antybody therapy.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) to the individual for applying the disclosure reduces Microglia activation in the individual.For example, in some embodiments, when its using one or more dosage as monotherapy or When being applied to the individual of disease or illness with complement-mediated with combination treatment, the anti-C1s antibody of theme reduces glue in the individual Cell plastid activation at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, until Few about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or More than 90%, compared to the microglia activation in the individual before the anti-C1s Antybody therapy.In some embodiments, The spongiocyte is astroglia or microglia.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) to the individual for applying the disclosure reduces Individual medium size lymphocyte infiltration.For example, in some embodiments, when its using one or more dosage as monotherapy or When being applied to the individual of disease or illness with complement-mediated with combination treatment, the anti-C1s antibody of theme reduces to be drenched in the individual Bar cellular infiltration at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, until Few about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or More than 90%, compared to the lymphocytic infiltration in the individual before the anti-C1s Antybody therapy.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) to the individual for applying the disclosure reduces Macrophages infiltration in the individual.For example, in some embodiments, when its using one or more dosage as monotherapy or When being applied to the individual of disease or illness with complement-mediated with combination treatment, the anti-C1s antibody of the disclosure (such as humanization Anti- C1s antibody) macrophages infiltration at least about 1%, at least about 5%, at least about 10%, at least about 15% in the individual are reduced, At least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, At least about 80%, at least about 90%, or more than 90%, compared to thin with the macrophage in the individual before the anti-C1s Antybody therapy Born of the same parents' infiltration.
In some embodiments, anti-C1s antibody (such as the anti-C1s antibody of humanization) to the individual for applying the disclosure reduces Antibody deposition in the individual.For example, in some embodiments, when it is using one or more dosage as monotherapy or with group Therapy is closed when being applied to the individual of disease or illness with complement-mediated, the anti-C1s antibody of the disclosure (such as humanization is anti- C1s antibody) antibody deposition at least about 1%, at least about 5%, at least about 10%, at least about 15% in the individual are reduced, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, compared to the antibody deposition in the individual before the anti-C1s Antybody therapy.
In some embodiments, apply the anti-C1s antibody of the disclosure to individual reduce anaphylatoxin in the individual (such as C3a, C4a, C5a) it generates.For example, in some embodiments, when it is using one or more dosage as monotherapy or with group Therapy is closed when being applied to the individual of disease or illness with complement-mediated, the anti-C1s antibody of the disclosure (such as humanization is anti- C1s antibody) anaphylatoxin generation at least about 1%, at least about 5%, at least about 10%, at least about 15% in the individual is reduced, until Few about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, until Few about 80%, at least about 90%, or more than 90%, compared to the anaphylatoxin in the individual before the anti-C1s Antybody therapy It generates horizontal.
The disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure or the anti-C1s comprising the disclosure resists The pharmaceutical compositions of body (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient are used to treat the disease with complement-mediated The purposes of the individual of disease or illness.In some embodiments, the disclosure provides anti-C1s antibody (such as the humanization of the disclosure Anti- C1s antibody) for treating the purposes of the individual of disease or illness with complement-mediated.In some embodiments, this public affairs It opens and the pharmaceutical composition of anti-C1s antibody (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient comprising the disclosure is provided Object is used to treat the purposes of the individual of disease or illness with complement-mediated.
The anti-C1s antibody (such as the anti-C1s antibody of humanization) that the disclosure provides the disclosure is mended in preparation for treating to have Purposes in the drug of the individual of disease or illness that body mediates.
The disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure or the anti-C1s comprising the disclosure resists The pharmaceutical compositions of body (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient are used to inhibit the purposes of complement activation. In some embodiments, the disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure or includes the disclosure Anti- C1s antibody (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient pharmaceutical compositions for inhibit have mend The purposes of complement activation in the individual of disease or illness that body mediates.In some embodiments, the disclosure provides the disclosure Anti- C1s antibody (such as the anti-C1s antibody of humanization) is used to inhibit the complement in the individual of the disease or illness with complement-mediated The purposes of activation.In some embodiments, the disclosure provides the anti-C1s antibody comprising the disclosure (such as the anti-C1s of humanization is anti- Body) and pharmaceutical acceptable excipient pharmaceutical compositions be used for inhibits have complement-mediated disease or illness individual in benefit The purposes of body activation.
The anti-C1s antibody (such as the anti-C1s antibody of humanization) that the disclosure provides the disclosure swashs in preparation for adjusting complement Purposes in drug living.In some embodiments, the Drug inhibition complement activation.In some embodiments, the drug Inhibit the complement activation for having in the disease of complement-mediated or the individual of illness.
The disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure or the anti-C1s comprising the disclosure resists The pharmaceutical compositions of body (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient are used for medical therapy.In some implementations In scheme, the anti-C1s antibody (such as the anti-C1s antibody of humanization) that the disclosure provides the disclosure is used for medical therapy.In some realities It applies in scheme, the disclosure provides anti-C1s antibody (such as the anti-C1s antibody of humanization) and pharmaceutically acceptable figuration comprising the disclosure The pharmaceutical compositions of agent are used for medical therapy.
The disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure or the anti-C1s comprising the disclosure resists The pharmaceutical compositions of body (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient are used to treat the disease with complement-mediated The individual of disease or illness.In some embodiments, the disclosure provides the anti-C1s antibody of the disclosure (such as the anti-C1s of humanization is anti- Body) for treating the individual of disease or illness with complement-mediated.In some embodiments, it includes this public affairs that the disclosure, which provides, The pharmaceutical compositions of the anti-C1s antibody (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient opened have for treating The disease of complement-mediated or the individual of illness.
The disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure or the anti-C1s comprising the disclosure resists The pharmaceutical compositions of body (such as the anti-C1s antibody of humanization) and pharmaceutical acceptable excipient are for adjusting complement activation.Some In embodiment, the disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) of the disclosure for adjusting complement activation. In some embodiments, the disclosure provides the anti-C1s antibody (such as the anti-C1s antibody of humanization) comprising the disclosure and pharmacy can Receive the pharmaceutical compositions of excipient for adjusting complement activation.In some embodiments, which inhibits complement to swash It is living.
The example of the non-limiting aspect of the disclosure
Above-described theme various aspects (including each embodiment) can be individually or with one or more other sides It is beneficial when face or combination of embodiment.It is not intended to limit the description of front, the certain non-of the disclosure of number 1-37 is provided below Restricted aspect.As that it is individual that each obviously can be used after reading the displosure to those skilled in the art The aspect of number is combined with any aforementioned or aftermentioned aspect numbered individually.This all such combination for being intended to various aspects mentions Combination for the various aspects for supporting and being not limited to hereafter clearly to provide:
The humanized antibody of the specific binding complement ingredient C1s of aspect 1. a kind of, wherein the antibody include:
A) heavy chain includes:
I) include amino acid sequence:
(Q/E)VQL(V/Q)QSGAE(V/L)KKPGASVK(L/V)SC(T/A)ASGFNIKDDYIHWV(K/R) QAPGQGLEWIGRIDPADGHTKYAPKFQVK(V/A)TITADTST(S/N)TAY(L/M)(E/Q)LSSL(R/T)SEDTAVY The area VH of YCARYGYGREVFDYWGQGTTVTVSS (SEQ ID NO:26)；With
Ii) comprising having the ammonia of at least 98% amino acid sequence identity with listed amino acid sequence in SEQ ID NO:28 The area Fc of base acid sequence, wherein amino acid 308 is Leu and amino acid 314 is Ser；With
B) light chain includes:
I) include amino acid sequence:
DIVLTQSPDSLAVSLGERATISCKASQSVDYDGDSYMNWYQQK(T/P)GQPPK(I/L)LIYDASNLES The VL of GIPARFSGSGSGTDFTLTISSLE (E/P) EDFA (I/V) YYCQQSNEDPWTFGGGTKVEIK (SEQ ID NO:27) Area；With
Ii) constant region of light chain.
The humanized antibody of 2. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:10；It and b) include SEQ ID The area VL of NO:20.
The humanized antibody of 3. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:10；It and b) include SEQ ID The area VL of NO:22.
The humanized antibody of 4. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:10；It and b) include SEQ ID The area VL of NO:24.
The humanized antibody of 5. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:12；It and b) include SEQ ID The area VL of NO:20.
The humanized antibody of 6. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:12；It and b) include SEQ ID The area VL of NO:22.
The humanized antibody of 7. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:12；It and b) include SEQ ID The area VL of NO:24.
The humanized antibody of 8. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:14；It and b) include SEQ ID The area VL of NO:20.
The humanized antibody of 9. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:14；It and b) include SEQ ID The area VL of NO:22.
The humanized antibody of 10. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:14；It and b) include SEQ ID The area VL of NO:24.
The humanized antibody of 11. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:16；It and b) include SEQ ID The area VL of NO:20.
The humanized antibody of 12. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:16；It and b) include SEQ ID The area VL of NO:22.
The humanized antibody of 13. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:16；It and b) include SEQ ID The area VL of NO:24.
The humanized antibody of 14. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:18；It and b) include SEQ ID The area VL of NO:20.
The humanized antibody of 15. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:18；It and b) include SEQ ID The area VL of NO:22.
The humanized antibody of 16. aspect 1 of aspect includes: a) including the area VH of SEQ ID NO:18；It and b) include SEQ ID The area VL of NO:24.
The humanized antibody of 17. aspect 1 of aspect, wherein the constant region of light chain is people's Kappa constant region of light chain.
The humanized antibody of 18. aspect 1 of aspect, wherein the heavy chain constant region includes listed amino acid in SEQ ID NO:28 Sequence.
The humanized antibody of 19. aspect 1 of aspect, wherein: a) heavy chain includes listed amino acid sequence in SEQ ID NO:29 Column；And b) light chain includes listed amino acid sequence in SEQ ID NO:30.
A kind of composition of aspect 20. includes: a) humanized antibody of any one of aspect 1-19；And b) pharmaceutically acceptable tax Shape agent.
A kind of container of aspect 21., it includes the compositions of aspect 20.
The container of 22. aspect 21 of aspect, wherein the container is sterile.
The container of 23. aspect 21 of aspect or aspect 22, wherein the container is phial, bottle, or syringe.
A kind of horizontal method for reducing the complement component cleaved products in individual of aspect 24., this method includes with effective Inhibit C1s and reduces antibody of the horizontal amount to any one of individual application aspect 1-19 of the cleaved products, or aspect 20 Composition.
The method of 25. aspect 24 of aspect, wherein the complement component cleaved products are C4 cleaved products.
The method of 26. aspect 25 of aspect, wherein the complement component cleaved products are C2 cleaved products.
The method of 27. aspect 25 of aspect, wherein the complement component cleaved products are C3 cleaved products.
The method of any one of 28. aspect 24-27 of aspect, wherein the individual is people.
Wantonly 8 one methods of 29. aspect 24-2 of aspect, wherein the application is intravenous.
The method of any one of 30. aspect 24-28 of aspect, wherein the application is intramuscular.
The method of any one of 31. aspect 24-28 of aspect, wherein the application is intrathecal.
The method of any one of 32. aspect 24-28 of aspect, wherein the application is subcutaneous.
The method of any one of 33. aspect 24-28 of aspect, wherein the illness of the reduction effectively treatment complement-mediated.
The method of 34. aspect 33 of aspect, wherein the illness of the complement-mediated is isoimmunization illness.
The method of 35. aspect 33 of aspect, wherein the illness of the complement-mediated is autoimmune disorder.
A kind of method for the cutting to complement component for inhibiting C1s to mediate in individual of aspect 36., this method includes to have The amount for the cutting to complement component that effect inhibits C1s to mediate applies the antibody of any one of aspect 1-19, or aspect 20 to the individual Composition.
A kind of method for the disease or illness that complement-mediated is treated in individual of aspect 37., this method includes effectively to control The amount of the disease or illness for the treatment of the complement-mediated applies the antibody of any one of aspect 1-19, or the combination of aspect 20 to the individual Object.
A kind of antibody of aspect 38. includes heavy chain and light chain, and wherein the heavy chain includes the area VH and heavy chain constant region, and this is light Chain includes the area VL；
Wherein the area VL includes VL CDR1, VL CDR2, and VL CDR3, and wherein the area VH includes VH CDR1, VH CDR2, and VH CDR3；
Wherein the VL CDR1 includes SEQ ID NO:1；
Wherein the VL CDR2 includes SEQ ID NO:2；
Wherein the VL CDR3 includes SEQ ID NO:3；
Wherein the VH CDR1 includes SEQ ID NO:4；
Wherein the VH CDR2 includes SEQ ID NO:5；
Wherein the VH CDR3 includes SEQ ID NO:6；
Wherein the heavy chain constant region includes IgG4 constant region, wherein the ammonia of the heavy chain constant region corresponding to SEQ ID NO:28 Base acid residue 308 is Leu, and the amino acid residue 314 of the heavy chain constant region corresponding to SEQ ID NO:28 is Ser；
And wherein the antibody specificity combines the C1s of activation.
The antibody of 39. aspect 38 of aspect, wherein the amino acid residue 108 of the heavy chain constant region corresponding to SEQ ID NO:28 It is Pro.
The antibody of 40. aspect 38 or 39 of aspect, wherein the amino acid of the heavy chain constant region corresponding to SEQ ID NO:28 is residual Base 115 is Glu.
A kind of antibody of aspect 41. includes heavy chain and light chain, and wherein the heavy chain includes the area VH and heavy chain constant region, and this is light Chain includes the area VL；
Wherein the area VL includes VL CDR1, VL CDR2, and VL CDR3, and wherein the area VH includes VH CDR1, VH CDR2, and VH CDR3；
Wherein the VL CDR1 includes SEQ ID NO:1；
Wherein the VL CDR2 includes SEQ ID NO:2；
Wherein the VL CDR3 includes SEQ ID NO:3；
Wherein the VH CDR1 includes SEQ ID NO:4；
Wherein the VH CDR2 includes SEQ ID NO:5；
Wherein the VH CDR3 includes SEQ ID NO:6；
Wherein the heavy chain constant region includes SEQ ID NO:28；
And wherein the antibody specificity combines the C1s of activation.
The antibody of any one of 42. aspect 38 to 41 of aspect, wherein the area VL includes and is selected from by SEQ ID NO:20,22, and The amino acid sequence of the group of 24 compositions.
The antibody of any one of 43. aspect 38 to 42 of aspect, wherein the area VH includes to be selected from by SEQ ID NO:10,12,14, 16, and 18 composition group amino acid sequence.
The antibody of any one of 44. aspect 38 to 43 of aspect, wherein:
(a) it includes SEQ ID NO:20 that the area VH, which includes the area NO:10, Qie Gai VL SEQ ID,；
(b) it includes SEQ ID NO:22 that the area VH, which includes the area NO:10, Qie Gai VL SEQ ID,；
(c) it includes SEQ ID NO:24 that the area VH, which includes the area NO:10, Qie Gai VL SEQ ID,；
(d) it includes SEQ ID NO:20 that the area VH, which includes the area NO:12, Qie Gai VL SEQ ID,；
(e) it includes SEQ ID NO:22 that the area VH, which includes the area NO:12, Qie Gai VL SEQ ID,；
(f) it includes SEQ ID NO:24 that the area VH, which includes the area NO:12, Qie Gai VL SEQ ID,；
(g) it includes SEQ ID NO:20 that the area VH, which includes the area NO:14, Qie Gai VL SEQ ID,；
(h) it includes SEQ ID NO:22 that the area VH, which includes the area NO:14, Qie Gai VL SEQ ID,；
(i) it includes SEQ ID NO:24 that the area VH, which includes the area NO:14, Qie Gai VL SEQ ID,；
(j) it includes SEQ ID NO:20 that the area VH, which includes the area NO:16, Qie Gai VL SEQ ID,；
(j) it includes SEQ ID NO:22 that the area VH, which includes the area NO:16, Qie Gai VL SEQ ID,；
(k) it includes SEQ ID NO:24 that the area VH, which includes the area NO:16, Qie Gai VL SEQ ID,；
(l) it includes SEQ ID NO:20 that the area VH, which includes the area NO:18, Qie Gai VL SEQ ID,；
(m) it includes SEQ ID NO:22 that the area VH, which includes the area NO:18, Qie Gai VL SEQ ID,；Or
(n) it includes SEQ ID NO:24 that the area VH, which includes the area NO:18, Qie Gai VL SEQ ID,.
The antibody of any one of 45. aspect 38 to 44 of aspect, wherein the area VH includes that the area SEQ ID NO:14, Qie Gai VL includes SEQ ID NO:22。
The antibody of any one of 46. aspect 38 to 45 of aspect, wherein the light chain further includes constant region of light chain.
The antibody of 47. aspect 46 of aspect, wherein the constant region of light chain includes SEQ ID NO:45.
The antibody of any one of 48. aspect 38 to 47 of aspect, wherein the heavy chain includes SEQ ID NO:29.
The antibody of any one of 49. aspect 38 to 48 of aspect, wherein the light chain includes SEQ ID NO:30.
The antibody of any one of 50. aspect 1 to 19 and 38 to 49 of aspect, is bispecific antibody or multi-specificity antibody.
A kind of immunoconjugates of the antibody comprising any one of aspect 1 to 19 and 38 to 50 of aspect 51..
A kind of nucleotide of the nucleotide set group of the antibody of any one of the encoding context 1 to 19 and 38 to 50 of aspect 52..
The aspect 53. a kind of carrier or carrier set group of the nucleotide of the nucleotide set group comprising aspect 52.
The place of a kind of nucleotide of nucleotide set group comprising aspect 52 of aspect 54. or the carrier of aspect 53 or carrier set group Chief cell.
A kind of antibody comprising any one of aspect 1 to 19 and 38 to 50 of aspect 55., the immunoconjugates of aspect 51, aspect 52 nucleotide or nucleotide set group, the carrier or carrier set group of aspect 53, or the host cell of aspect 54, and it is pharmaceutically acceptable The pharmaceutical compositions of excipient.
A kind of method for inhibiting complement pathway in subject in need of aspect 56., comprising applying medicine to the subject Learn the antibody of any one of a effective amount of aspect 1 to 19 and 38 to 50, the immunoconjugates of aspect 51, the nucleotide or core of aspect 52 Thuja acid set group, the carrier or carrier set group, the host cell of aspect 54, or the pharmaceutical compositions of aspect 55 of aspect 53.
A kind of method for the cutting to complement component C4 for inhibiting C1s to mediate in subject in need of aspect 57., packet Antibody containing any one of the aspect 1 to 19 and 38 to 50 for applying pharmacy effective dose to the subject, the immunoconjugates of aspect 51, The nucleotide or nucleotide set group of aspect 52, the carrier of aspect 53 or carrier set group, the host cell of aspect 54, or aspect 55 Pharmaceutical compositions.
A kind of method for the disease or illness that complement-mediated is treated in subject in need of aspect 58., comprising to this Subject applies the antibody of any one of aspect 1 to 19 and 38 to 50 of pharmacy effective dose, the immunoconjugates of aspect 51, aspect 52 Nucleotide or nucleotide set group, the carrier of aspect 53 or the pharmacy group of carrier set group, the host cell of aspect 54, or aspect 55 Close object.
The method of 59. aspect 58 of aspect, wherein the disease of the complement-mediated or illness are selected from by age related macular degeneration, Alzheimer (Alzheimer) family name disease, amyotrophic lateral sclerosis, allergic reaction, argyrophilic grain dementia, arthritis (such as Rheumatoid arthritis), asthma, atherosclerosis, atypia hemolytic uremic syndrome, autoimmune disease (including Such as autoimmune hemolytic anemia (AIHA)；Tepid-type AIHA；Mixed type AIHA；Deng), Braak that-simon this (Barraquer-Simons) syndrome, Bei QieteFamily name's disease, Britain's type amyloid angiopathy, epidermolysis class day blister Sore, Burger (Buerger) family name disease, C1q nephrosis, cancer, catastrophic (catastrophic) antiphospholipid syndrome, brain amyloid blood Pipe disease, cold coagulation disease, corticobasal degeneration, Creutz Fei Erte-Jacob (Creutzfeldt-Jakob) disease, Crow Grace (Crohn) family name disease, cryoglobulinemic vasculitis, boxer's dementia, dementia with Lewy body (DLB), with diffusing for calcification Nerve fibrinogen tangles, lupus erythematosus discoides, Tang (Down) Cotard, Ai Wen (Evan) Cotard, focal segment Property glomerulosclerosis, formal thought disorder, Frontotemporal dementia (FTD), with the chain volume temporo with Parkinson's disease of chromosome 17 Leaf is dull-witted, frontotemporal lobar degeneration, Gerstmann-Straussler-Scheinker disease, guillain-Barre (Guillain-Barr é) Syndrome, Halle Wo Deng-Shi Paci (Hallervorden-Spatz) disease, hemolytic uremic syndrome, inherited vascular water It is swollen, hypophosphatemia, idiopathic pneumonia syndrome, immune complex disease, inclusion body myositis, communicable disease (such as by thin Bacterium (such as Neisseria meningitidis or streptococcus), viral (such as human immunodeficiency virus (HIV)), or other infectious agents draw The disease risen), diseases associated with inflammation, ischemia/reperfusion injury, mild cognitive impairment, immunologic thrombocytopenic purpura (ITP), A type deficiency of molybdenum cofactor disease (MoCD), I type membrano proliferative glomerulonephritis (MPGN), II type Membrane proliferative glomerulus Ephritis (MPGN) (compactness storage disorders), membraneous nephritis, multiple infarct dementia, lupus (such as systemic loupus erythematosus (SLE)), glomerulonephritis, Kawasaki (Kawasaki) disease, multifocal motor neuropathy, multiple sclerosis, multi-system atrophy, weight Disease myasthenia, myocardial infarction, myotonia dystrophy, neuromyelitis optica, c-type Niemann-pik (Niemann-Pick) disease, With non-Guam type motor neuron disease of neurofibrillary tangles, Parkinson (Parkinson) family name disease, with dull-witted pa gold Sen Shi disease, paroxysmal nocturnal hemoglobinuria, pemphigus vulgaris, pik (Pick) family name disease, postencephalitic parkinsonism, polymyarian Inflammation, prion protein cerebral amyloid angiopathy, gliosis under progressive cortex, stein-leventhal syndrome, silver bits Disease, septicemia, shiga toxin Escherichia coli (STEC)-HuS, Duchenne-Arandisease, apoplexy, subacute sclerosing panencephalitis, only Entanglement type is dull-witted, graft rejection, vasculitis (such as ANCA relevant blood vessel is scorching), Wegener (Wegner) family name's granulomatosis, sickle Shape cytopathy, cryoglobulinemia, mixed type cryoglobulinemia, idiopathic (essential) mixed type cryoglobulinemia, II type mixed type cryoglobulinemia, type III mixed type cryoglobulinemia, ephritis, drug-induced thrombopenia, wolf Sore ephritis, acquired epidermolysis bullosa, delaying type hemolytic blood transfusion reaction, hypocomplementemia nettle rash vasculitis are comprehensive Simulator sickness, false bullous keratopathy, blood platelet refractoriness, chronic inflammation demyelinating polyneuropathy (CIDP), myelosis Abnormal syndrome (MDS), Miller-fischer (Miller-Fisher) syndrome, acute inflammatory demyelinating polyneuropathy (AIDP), acute exercise axon nerve is sick (AMAN), and acute exercise and feeling axon nerve are sick (AMSAN), pharynx-neck-arm Variant, and the group of any combination thereof composition.
The method of 60. aspect 58 or 59 of aspect, wherein the disease of the complement-mediated or illness are selected from by epidermolysis class day blister Sore, cold coagulation disease, autoimmune hemolytic anemia (AIHA), immunologic thrombocytopenic purpura (ITP), multifocal fortune Dynamic neuropathy, neuromyelitis optica, and the group of any combination thereof composition.
The method of any one of 61. aspect 56 to 60 of aspect, wherein parenterally, intravenously, subcutaneously, intradermal, transdermal, muscle It is interior, it takes orally, intraocularly, intrathecal, in peritonaeum, intranasally, oral cavity is sublingual, rectum, vagina, or applies the antibody via lung path.
Embodiment
The following example is proposed to provide how to manufacture and use the complete of the disclosure to those of ordinary skill in the art Disclosure and description, and following description is not intended to be limited to inventors believe that it is scope of the disclosure, is not intended to expression Experiment is the experiment for the progress completely or only having below.It has tried to make sure that about used number (for example, amount, temperature etc.) Accuracy, but cope with some experimental errors and deviation is illustrated.Unless otherwise stated, number is parts by weight, point Son amount is weight average molecular weight, and temperature is degree Celsius and pressure is atmospheric pressure or close to atmospheric pressure.Standardized abbreviations can be used, Such as bp, base-pair；Kb, kilobase；Pl, picoliters；S or sec, second；Min, minute；H or hr, hour；Aa, amino acid；Kb, thousand Base；Bp, base-pair；Nt, nucleotide；I.m., intramuscular (ground)；I.p., (ground) in peritonaeum；S.c., subcutaneous (ly)； Deng.
Embodiment 1: the anti-aC1s variant of humanization
Produce the humanization variants of anti-aC1s.Additionally provide the amino acid sequence in the domain heavy chain VH of humanization variants 1-5； Encode the nucleotide sequence in the domain heavy chain VH of the humanization variants.The light chain VL of humanization variants 1,2 and 5 is shown in Fig. 6-8 The amino acid sequence in domain, and encode the nucleotide sequence in the domain light chain VL of the humanization variants.Amino relative to the anti-aC1s of mouse Acid sequence (VL SEQ ID NO:7；VH SEQ ID NO:8) amino acid of differences summarize in figures 9 and 10.
Single letter amino acid code is following (being indicated with the 3- letter amino acid code in bracket):
G- glycine (Gly)
P- proline (Pro)
A- alanine (Ala)
V- valine (Val)
L-Leu (Leu)
I- isoleucine (Ile)
M- methionine (Met)
C- cysteine (Cys)
F- phenylalanine (Phe)
Y- tyrosine (Tyr)
W- tryptophan (Trp)
H- histidine (His)
K- lysine (Lys)
R- arginine (Arg)
Q- glutamine (Gln)
N- asparagine (Asn)
E- glutamic acid (Glu)
D-Asp (Asp)
S- serine (Ser)
T- tryptophan (Thr)
Embodiment 2: the characterization of the anti-aC1s variant of humanization
The binding characteristic of the anti-aC1s variant of humanization provides (being Figure 11 and Figure 12 respectively) in table 4 and 5.For various people The anti-aC1s variant of sourceization provides (the first field) to the relative binding affinity of the C1s of activation in table 4, and the table 4 is in Figure 11 In show.
Produce all 15 compositions (VH variant 1+Vk variant 1；VH variant 1+Vk variant 2；VH variant 1+Vk variant 5； VH variant 2+Vk variant 1；VH variant 2+Vk variant 2；VH variant 2+Vk variant 5；VH variant 3+Vk variant 1；VH variant 3+Vk becomes Body 2；VH variant 3+Vk variant 5；VH variant 4+Vk variant 1；VH variant 4+Vk variant 2；VH variant 4+Vk variant 5；VH variant 5+ Vk variant 1；VH variant 5+Vk variant 2；VH variant 5+Vk variant 5).It tests each humanization variants and biotinylated mouse is anti- The ability of aC1s competitive binding activity C1s.The data are shown in Figure 11, the second field.
Each humanization variants are tested in the commercially available measuring method of measurement classical pathway of complement (CP) activation.The result is being schemed 11, it shows in third field.The data show that all 15 humanization variants inhibit CP activation, IC50With the anti-aC1s's of mouse IC50It is similar.
The Kinetic Characterization of affinity is combined to the anti-aC1s variant of 8 kinds of humanizations.The data describe in table 5, should Table 5 is shown in FIG. 12.
Embodiment 3: the In vivo study in machin
In order to assess the pharmacokinetics (PK) and pharmacodynamics (PD) characteristic of the anti-aC1s of humanization, in machin (Macaca Fascicularis the single dose quantity research of the anti-aC1s of humanization is carried out in).In addition, in order to compare the people by various administration method The bioavilability of the anti-aC1s of sourceization, the anti-aC1s variant of the humanization pass through intravenous (IV) or subcutaneous (SC) injection application.It gives After the anti-aC1s of medicine humanization, blood plasma and blood serum sample are acquired at the specified time point to measure the circulation composition of the anti-aC1s of humanization； And inhibition of the anti-aC1s of humanization to classic complement approach (CP) is passed through with assessment.The blood plasma of the anti-aC1s of humanization at any time and Serum levels provide PK data；CP at any time inhibits to provide PD data.
All research animals are females, and weight is between 2.4-3.9kg, and the age is between 3-5 years old.In addition, all Animal did not received drug administration.
Whole blood is collected in K2In EDTA pipe and serum separator pipe, it is respectively used to blood plasma and serum processing, and store immediately At -15 DEG C to -25 DEG C.
In order to assess the anti-aC1s variant VH3/VK2-Fc-sub of humanization4Pharmacokinetic profile, will in figs. 13 and 14 The plasma sample of the time point acquisition of description is diluted and runs in ELISA with quantitative VH3/VK2-Fc-sub4Blood plasma is dense Degree.In short, diluted plasma sample to be added to 96 orifice plates pre-coated with the C1s of activation.Plasma sample be incubated for and with After washing afterwards, the detection antibody of the conjugation horseradish peroxidase special to human IgG is added to detect the VH3/ for combining C1s VK2-Fc-sub4.Finally, 3,3 ', 5,5 '-tetramethyl benzidine (TMB) substrates of addition are read on spectrophotometer with originating Chrominance response.By from the VH3/VK2-Fc-sub parallel with plasma sample4Standard curve interpolation, determine all samples The VH3/VK2-Fc-sub of product4Plasma concentration.
It usesClassic complement approach kit assesses VH3/VK2-Fc-sub4Pharmacodynamic effects.It shouldKit is commercially available, and is related to the purposes of enzyme-linked immunosorbent assay (ELISA), is intended to pass through work Change the classical pathway of vitro samples and measures the in vitro generation of the final decomposition product (C5b-9) of the approach to assess blood serum sample In the active intensity of classic complement approach.Sample is measured according to the specification of manufacturer.In short, by the serum from monkey Sample (its time point indicated in figures 14 and 15 is collected) is diluted and is added to the hole of provided 96 orifice plates In.After incubation, the special detection antibody of final decomposition product (C5b-9) of the addition to classical pathway, and on spectrophotometer Measure the chrominance response.The all samples of single monkey are compared and be standardized into sample before the administration of same monkey (to Before medicine=100% activity).
Figure 13, which describes, comes from 3 administration VH3/VK2-Fc-sub4The PD and PK of the animal of (intravenously being applied with 10mg/kg) Data.As shown in Figure 13, within the period up to 650 hours (27 days), VH3/VK2-Fc-sub4Serum-concentration 70 Between μ g/mL and 300 μ g/mL.In this same time period, inhibit CP activity up to 80% to 99%.
Figure 14, which describes, comes from 3 administration VH3/VK2-Fc-sub4PD the and PK number of the animal of (with 20mg/kg subcutaneous administration) According to.As shown in Figure 14, within the long period for reaching 85 days, VH3/VK2-Fc-sub4Serum-concentration in about 50 μ g/mL and Between about 450 μ g/mL.In this same time period, inhibit as usedThe CP activity of kits reaches 60% to 99%.Further, the VH3/VK2-Fc-sub of single 20mg/kg subcutaneous dosage4Complement pathway is inhibited to be greater than 90% Up to 28 days (Figure 19).
Embodiment 4: the characterization of the anti-C1s antibody comprising the modified area Fc
The anti-aC1s variant (VH3/VK2) of the humanization includes human IgG 4, which there is S241P and L248E to replace.In order to Enhance half-life period and subcutaneous bioavilability, modifies the area Fc of VH3/VK2 to include that M428L and N434S replaces.Obtained by it was found that Antibody (referred to as VH3/VK2-Fc-sub4) with 1.53x10-9Dissociation constant specific binding activity C1s.
VH3/VK2-Fc-sub4Inhibit in vitro complement pathway to conventional anti-C1s antibody (previously in U.S. Patent number 8, It is described as VH4/VK2 in 945,562) similar degree.In vitro in classic complement approach serum active measuring method, VH3/VK2- Fc-sub4(circle) display and anti-C1s (box) similar ED50Although passing through VH3/VK2-Fc-sub4Inhibit classical complement way Diameter activity is than inhibiting complement activity more slowly (Figure 17 A) by anti-C1s.In addition, VH3/VK2-Fc-sub4Display resists with conventional The similar hemolysis levels of C1s antibody (Figure 17 B).
Assuming that if FcRn, which is combined, is related to the VH3/VK2 (VH3/VK2-Fc-sub that the FcRn without enhancing is combined4) Recycling, then VH3/VK2-Fc-sub4Should have longer half-life period and therefore extended pharmacodynamic effects, compared to VH3/ VK2.The VH3/VK2 or VH3/VK2-Fc-sub of the 10mg/kg of single intravenous dosages are applied to machin4, and periodically extract Blood.After application in 650 hours, VH3/VK2-Fc-sub applied4Animal than apply VH3/VK2 animal in its blood In there is the level (Figure 18 A) of the higher antibody always.In addition, VH3/VK2-Fc-sub4With medicine more extended than VH3/VK2 Effect learns effect.The 10mg/kg VH3/VK2-Fc-sub of single dosage4Inhibition complement pathway activity is more than in 650 hours 70%, however the VH3/VK2 of same dose display inhibits to be gradually reduced, and almost immediately begins to, and almost reach 650 small after application When intermedium control level (Figure 18 B).Also in the 20mg/kg VH3/VK2-Fc-sub that applied single subcutaneous dosage4's These effects are observed in machin (referring to Figure 14).
Embodiment 5: the pharmacokinetic studies of the anti-C1s antibody in machin medium sized vein and after subcutaneous administration
The purpose of this Inquiry research is assessment VH3/VK2-Fc-sub4Pharmacokinetics, (IV) is injected in single vein Afterwards, subcutaneous (SC) injection once a week after single IV is injected in female cynomolgus monkeys, or repeat SC injection.
Researching and designing
Animal is distributed to each group and is treated as shown in table 2.To use the butterfly infusion line that is pre-installed via In peripheral vein medium sized vein (IV) inject or pass through back omoplate area in subcutaneous (SC) inject administration animal.If in injection part Stimulation is noticed in position, then pectus region can be used for subsequent SC injection to avoid further stimulation.The administration frequency and examination The expection pharmacokinetics for testing product is consistent.It is expected that studying the therapeutic scheme of selection for this to identify accessible concentration in peripheral blood With relevant pharmacological activity.
Table 2: distribution group
aTotal administered volume (mL) will be calculated based on nearest weight.
CA/V: control product/medium；IV: intravenous push；SC: subcutaneous bolus injection.
Clinical observation
Before cleaning indoors, clinical observation will carry out once daily, start from every animal adapts in AM second day. Death rate inspection will carry out twice daily, to assess general animal health and situation (wellness).
When necessary, additional clinical observation can be carried out.It, will be into if the animal patient's condition of the clinical observation display decline of animal Row animal doctor assesses and notifies head of research.
It will be from controlled, the peripheral vein collection blood of conscious animal.To after administration first 24 hours, Blood will not be collected from the vein (or limbs) for IV administration application.It is extracted via single and collects blood, then suitably Ground separates.
In the case where non-periodically ptomatopsia, if it would be possible, will be from conscious dying animal in its anesthesia Venous samples are collected before.
Will following time point collect blood sample, and be stored in front for the treatment of it is wet on ice:
1st and 3 group: the 1st day (15 minutes after administration, 30 minutes, 1 hour, 2 hours and 4 hours), the 2nd day (after administration 24 hours), the 5th day, the 8th day (before administration, 30 minutes, 1 hour, 2 hours and 4 hours after administration), and the 9th day, the 10th day, the 11 days, the 12nd day, the 13rd day, the 14th day, the 15th day (before administration), the 18th day, the 22nd day (before administration), the 25th day, the 29th day (before administration), the 32nd day, the 36th day (before administration), the 39th day, the 43rd day (before administration), the 46th day, the 50th day, the 53rd day, and 57th day；
2nd group: the 1st day (after administration 15 minutes, 30 minutes, 1 hour, 2 hours and 4 hours), (24 is small after administration within the 2nd day When), the 3rd day (after administration 48 hours), the 5th day (after administration 96 hours), the 8th day (after administration 168 hours), the 15th day, the 22nd It, the 25th day, the 29th day, the 32nd day, the 36th day, the 39th day, the 43rd day, the 46th day, the 50th day, the 53rd day and the 57th day；With
4th group: the 1st day (after administration 30 minutes, 1 hour, 2 hours and 4 hours), the 2nd day (after administration 24 hours), the 3rd It, the 4th day, the 5th day, the 6th day, the 11st day, the 15th day, the 18th day, the 22nd day, the 25th day, the 29th day (before administration, after administration 30 minutes, 1 hour, 2 hours and 4 hours), the 30th day, the 31st day, the 32nd day, the 33rd day, the 34th day, the 39th day, the 43rd day, 46th day, the 50th day, the 53rd day and the 57th day.
Although describing the disclosure by reference to the specific embodiment of the disclosure, those skilled in the art should be managed Solution, in the case where not departing from the true spirit and range of the disclosure, can carry out various changes, and alternative equivalent.In addition, Can carry out it is many change so that specific condition, material, material composition, process, processing step is applicable in the purpose of the disclosure, spirit with Range.All such modifications are intended in the scope of the appended claims.
Sequence table
<110>U.S.'s ratio Ao Weiladi Wei Stocks Trading Co Co., Ltd (BIOVERATIV USA INC.)
S Pa Nike (PANICKER, SANDIP)
G pa is auspicious (PARRU, GRAHAM)
NE Si Tangelianao (STAGLIANO, NANCY E.)
<120>anti-C1s antibody and its application method
<130> 4159.504PC01/C-K/BMD
<150> US 62/407,390
<151> 2016-10-12
<160> 54
<170> PatentIn version 3.5
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<213>artificial sequence
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<223> VL CDR1
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Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn
1 5 10 15
<210> 2
<211> 7
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<213>artificial sequence
<220>
<223> VL CDR2
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Asp Ala Ser Asn Leu Glu Ser
1 5
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<211> 9
<212> PRT
<213>artificial sequence
<220>
<223> VL CDR3
<400> 3
Gln Gln Ser Asn Glu Asp Pro Trp Thr
1 5
<210> 4
<211> 5
<212> PRT
<213>artificial sequence
<220>
<223> VH CDR1
<400> 4
Asp Asp Tyr Ile His
1 5
<210> 5
<211> 17
<212> PRT
<213>artificial sequence
<220>
<223> VH CDR2
<400> 5
Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe Gln
1 5 10 15
Val
<210> 6
<211> 10
<212> PRT
<213>artificial sequence
<220>
<223> VH CDR3
<400> 6
Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr
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<210> 7
<211> 111
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<213>artificial sequence
<220>
<223>the anti-C1s VL of parent mouse can lighten
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Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Thr Gly Gln Pro Pro
35 40 45
Lys Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His
65 70 75 80
Pro Val Glu Glu Glu Asp Ala Ala Ile Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
<210> 8
<211> 119
<212> PRT
<213>artificial sequence
<220>
<223>the VH CDR of anti-C1s antibody
<400> 8
Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Leu Thr Val Ser Ser
115
<210> 9
<211> 673
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<220>
<223>people C1s
<400> 9
Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala
1 5 10 15
Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly
20 25 30
Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu
35 40 45
Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu
50 55 60
Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile
65 70 75 80
Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys
85 90 95
Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr
100 105 110
Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys
115 120 125
Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro
130 135 140
Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys
145 150 155 160
Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn
165 170 175
Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg
180 185 190
Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe
195 200 205
Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val
210 215 220
Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe
225 230 235 240
Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile
245 250 255
Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr
260 265 270
His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val
275 280 285
Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile
290 295 300
Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr
305 310 315 320
Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys
325 330 335
Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn
340 345 350
Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg
355 360 365
Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly
370 375 380
Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly
385 390 395 400
Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro
405 410 415
Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys
420 425 430
Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala
435 440 445
Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly
450 455 460
Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser
465 470 475 480
Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His
485 490 495
Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp
500 505 510
Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro
515 520 525
Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu
530 535 540
Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys
545 550 555 560
Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro
565 570 575
Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala
580 585 590
Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys
595 600 605
Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln
610 615 620
Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp
625 630 635 640
Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr
645 650 655
Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu
660 665 670
Asp
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<400> 10
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Leu Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Ala Thr Ile Thr Ala Asp Thr Ser Thr Asn Thr Ala Tyr
65 70 75 80
Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser
115
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<212> DNA
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<223>VH variant 1
<400> 11
gaggttcagc tggtgcagtc tggggctgag cttaagaagc caggggcctc agtcaagttg 60
tcctgcacag cttctggctt taacattaaa gacgactata tacactgggt gaagcaggcc 120
cctggacagg gcctggagtg gattggaagg attgatcctg cggatggtca tactaaatat 180
gccccgaagt tccaagtcaa ggccactata actgcagaca catccaccaa cacagcctac 240
ctgcagctca gcagcctgac atctgaggac actgccgtct attactgtgc tagatatggt 300
tacgggaggg aggtctttga ctactggggc caaggcacca ctgtcacagt ctcctca 357
<210> 12
<211> 119
<212> PRT
<213>artificial sequence
<220>
<223>VH2 variant
<400> 12
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Ala Thr Ile Thr Ala Asp Thr Ser Thr Asn Thr Ala Tyr
65 70 75 80
Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser
115
<210> 13
<211> 357
<212> DNA
<213>artificial sequence
<220>
<223>VH variant 2
<400> 13
gaggttcagc tggtgcagtc tggggctgag gtgaagaagc caggggcctc agtcaagttg 60
tcctgcacag cttctggctt taacattaaa gacgactata tacactgggt gaagcaggcc 120
cctggacagg gcctggagtg gattggaagg attgatcctg cggatggtca tactaaatat 180
gccccgaagt tccaagtcaa ggccactata actgcagaca catccaccaa cacagcctac 240
ctggagctca gcagcctgag atctgaggac actgccgtct attactgtgc tagatatggt 300
tacgggaggg aggtctttga ctactggggc caaggcacca ctgtcacagt ctcctca 357
<210> 14
<211> 119
<212> PRT
<213>artificial sequence
<220>
<223>VH3 variant
<400> 14
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser
115
<210> 15
<211> 357
<212> DNA
<213>artificial sequence
<220>
<223>VH variant 3
<400> 15
caggttcagc tggtgcagtc tggggctgag gtgaagaagc caggggcctc agtcaagttg 60
tcctgcacag cttctggctt taacattaaa gacgactata tacactgggt gaagcaggcc 120
cctggacagg gcctggagtg gattggaagg attgatcctg cggatggtca tactaaatat 180
gccccgaagt tccaagtcaa agtcactata actgcagaca catccaccag cacagcctac 240
ctggagctca gcagcctgag atctgaggac actgccgtct attactgtgc tagatatggt 300
tacgggaggg aggtctttga ctactggggc caaggcacca ctgtcacagt ctcctca 357
<210> 16
<211> 119
<212> PRT
<213>artificial sequence
<220>
<223>VH4 variant
<400> 16
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser
115
<210> 17
<211> 357
<212> DNA
<213>artificial sequence
<220>
<223>VH variant 4
<400> 17
caggttcagc tggtgcagtc tggggctgag gtgaagaagc caggggcctc agtcaaggtc 60
tcctgcacag cttctggctt taacattaaa gacgactata tacactgggt gcgccaggcc 120
cctggacagg gcctggagtg gattggaagg attgatcctg cggatggtca tactaaatat 180
gccccgaagt tccaagtcaa agtcactata actgcagaca catccaccag cacagcctac 240
atggagctca gcagcctgag atctgaggac actgccgtct attactgtgc tagatatggt 300
tacgggaggg aggtctttga ctactggggc caaggcacca ctgtcacagt ctcctca 357
<210> 18
<211> 119
<212> PRT
<213>artificial sequence
<220>
<223>VH5 variant
<400> 18
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser
115
<210> 19
<211> 357
<212> DNA
<213>artificial sequence
<220>
<223>VH variant 5
<400> 19
caggttcagc tggtgcagtc tggggctgag gtgaagaagc caggggcctc agtcaaggtc 60
tcctgcgcag cttctggctt taacattaaa gacgactata tacactgggt gcgccaggcc 120
cctggacagg gcctggagtg gattggaagg attgatcctg cggatggtca tactaaatat 180
gccccgaagt tccaagtcaa agtcactata actgcagaca catccaccag cacagcctac 240
atggagctca gcagcctgag atctgaggac actgccgtct attactgtgc tagatatggt 300
tacgggaggg aggtctttga ctactggggc caaggcacca ctgtcacagt ctcctca 357
<210> 20
<211> 111
<212> PRT
<213>artificial sequence
<220>
<223>1 variant of V Kappa can lighten
<400> 20
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Thr Gly Gln Pro Pro
35 40 45
Lys Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Glu Glu Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105 110
<210> 21
<211> 333
<212> DNA
<213>artificial sequence
<220>
<223>V Kappa variant 1
<400> 21
gacattgtgc tgacccaatc tccagactct ttggctgtgt ctctcgggga gagggccacc 60
atctcctgca aggccagcca aagtgttgat tatgatggtg atagttatat gaactggtac 120
caacaaaaaa caggacagcc acccaaaatc ctcatttatg atgcatccaa tttggaatct 180
ggcatcccag ccaggtttag tggcagtggg tctgggacag acttcaccct caccatcagc 240
agcctggagg aggaggattt tgcaatctat tactgtcagc aaagtaatga agacccgtgg 300
acgttcggtg gaggcaccaa ggtggaaatc aaa 333
<210> 22
<211> 111
<212> PRT
<213>artificial sequence
<220>
<223>2 variant of V Kappa can lighten
<400> 22
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Glu Pro Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105 110
<210> 23
<211> 333
<212> DNA
<213>artificial sequence
<220>
<223>V Kappa variant 2
<400> 23
gacattgtgc tgacccaatc tccagactct ttggctgtgt ctctcgggga gagggccacc 60
atctcctgca aggccagcca aagtgttgat tatgatggtg atagttatat gaactggtac 120
caacaaaaac caggacagcc acccaaaatc ctcatttatg atgcatccaa tttggaatct 180
ggcatcccag ccaggtttag tggcagtggg tctgggacag acttcaccct caccatcagc 240
agcctggagc ctgaggattt tgcaatctat tactgtcagc aaagtaatga agacccgtgg 300
acgttcggtg gaggcaccaa ggtggaaatc aaa 333
<210> 24
<211> 111
<212> PRT
<213>artificial sequence
<220>
<223>5 variant of V Kappa can lighten
<400> 24
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105 110
<210> 25
<211> 333
<212> DNA
<213>artificial sequence
<220>
<223>V Kappa variant 5
<400> 25
gacattgtgc tgacccaatc tccagactct ttggctgtgt ctctcgggga gagggccacc 60
atctcctgca aggccagcca aagtgttgat tatgatggtg atagttatat gaactggtac 120
caacaaaaac caggacagcc acccaaactc ctcatttatg atgcatccaa tttggaatct 180
ggcatcccag ccaggtttag tggcagtggg tctgggacag acttcaccct caccatcagc 240
agcctggagc ctgaggattt tgcagtctat tactgtcagc aaagtaatga agacccgtgg 300
acgttcggtg gaggcaccaa ggtggaaatc aaa 333
<210> 26
<211> 119
<212> PRT
<213>artificial sequence
<220>
<223>area VH of anti-C1s antibody
<220>
<221>hybrid characteristic
<222> (1)..(1)
<223>it can be Gln or Glu
<220>
<221>hybrid characteristic
<222> (5)..(5)
<223>it can be Val or Gln
<220>
<221>hybrid characteristic
<222> (11)..(11)
<223>it can be Val or Leu
<220>
<221>hybrid characteristic
<222> (20)..(20)
<223>it can be Leu or Val
<220>
<221>hybrid characteristic
<222> (23)..(23)
<223>it can be Thr or Ala
<220>
<221>hybrid characteristic
<222> (38)..(38)
<223>it can be Lys or Arg
<220>
<221>hybrid characteristic
<222> (68)..(68)
<223>it can be Val or Ala
<220>
<221>hybrid characteristic
<222> (77)..(77)
<223>it can be Ser or Asn
<220>
<221>hybrid characteristic
<222> (81)..(81)
<223>it can be Leu or Met
<220>
<221>hybrid characteristic
<222> (82)..(82)
<223>it can be Glu or Gln
<220>
<221>hybrid characteristic
<222> (87)..(87)
<223>it can be Arg or Thr
<400> 26
Xaa Val Gln Leu Xaa Gln Ser Gly Ala Glu Xaa Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Xaa Ser Cys Xaa Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Xaa Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Xaa Thr Ile Thr Ala Asp Thr Ser Thr Xaa Thr Ala Tyr
65 70 75 80
Xaa Xaa Leu Ser Ser Leu Xaa Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser
115
<210> 27
<211> 111
<212> PRT
<213>artificial sequence
<220>
<223>area VL of anti-C1s antibody
<220>
<221>hybrid characteristic
<222> (44)..(44)
<223>it can be Thr or Pro
<220>
<221>hybrid characteristic
<222> (50)..(50)
<223>it can be Ile or Leu
<220>
<221>hybrid characteristic
<222> (84)..(84)
<223>it can be Glu or Pro
<220>
<221>hybrid characteristic
<222> (89)..(89)
<223>it can be Ile or Val
<400> 27
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Xaa Gly Gln Pro Pro
35 40 45
Lys Xaa Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Glu Xaa Glu Asp Phe Ala Xaa Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105 110
<210> 28
<211> 327
<212> PRT
<213>artificial sequence
<220>
<223>4 constant region of human IgG (Fc) variant 2
<400> 28
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
1 5 10 15
Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr
65 70 75 80
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro
100 105 110
Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
115 120 125
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
130 135 140
Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
145 150 155 160
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
180 185 190
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu
195 200 205
Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
210 215 220
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys
225 230 235 240
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
245 250 255
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
260 265 270
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
275 280 285
Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
290 295 300
Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln Lys Ser
305 310 315 320
Leu Ser Leu Ser Leu Gly Lys
325
<210> 29
<211> 446
<212> PRT
<213>artificial sequence
<220>
<223>VH3 maturation heavy chain
<400> 29
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val
260 265 270
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His
420 425 430
Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
435 440 445
<210> 30
<211> 218
<212> PRT
<213>artificial sequence
<220>
<223>2 variant maturation light chain of V Kappa
<400> 30
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Glu Pro Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105 110
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
115 120 125
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
145 150 155 160
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
195 200 205
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 31
<211> 465
<212> PRT
<213>artificial sequence
<220>
<223>synthetic amino acid array
<400> 31
Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg
1 5 10 15
Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
20 25 30
Pro Gly Ala Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile
35 40 45
Lys Asp Asp Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu
50 55 60
Glu Trp Ile Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala
65 70 75 80
Pro Lys Phe Gln Val Lys Val Thr Ile Thr Ala Asp Thr Ser Thr Ser
85 90 95
Thr Ala Tyr Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
100 105 110
Tyr Tyr Cys Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp
115 120 125
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
130 135 140
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr
145 150 155 160
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
165 170 175
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
180 185 190
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
195 200 205
Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp
210 215 220
His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr
225 230 235 240
Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro
245 250 255
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
260 265 270
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp
275 280 285
Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
290 295 300
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val
305 310 315 320
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
325 330 335
Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys
340 345 350
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
355 360 365
Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
370 375 380
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
385 390 395 400
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
405 410 415
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys
420 425 430
Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu His Glu
435 440 445
Ala Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
450 455 460
Lys
465
<210> 32
<211> 238
<212> PRT
<213>artificial sequence
<220>
<223>synthetic amino acid array
<400> 32
Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Gly Ala Arg Cys Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala
20 25 30
Val Ser Leu Gly Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser
35 40 45
Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro
50 55 60
Gly Gln Pro Pro Lys Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser
65 70 75 80
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Ile Tyr Tyr Cys
100 105 110
Gln Gln Ser Asn Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val
115 120 125
Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro
130 135 140
Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu
145 150 155 160
Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175
Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser
180 185 190
Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala
195 200 205
Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
210 215 220
Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 33
<211> 11
<212> PRT
<213>artificial sequence
<220>
<223>the VL CDR1 (CDR-L1) of anti-C1s antibody
<400> 33
Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr
1 5 10
<210> 34
<211> 3
<212> PRT
<213>artificial sequence
<220>
<223>the VL CDR2 (CDR-L2) of anti-C1s antibody
<400> 34
Asp Ala Ser
1
<210> 35
<211> 6
<212> PRT
<213>artificial sequence
<220>
<223>the VL CDR3 (CDR-L3) of anti-C1s antibody
<400> 35
Ser Asn Glu Asp Pro Trp
1 5
<210> 36
<211> 7
<212> PRT
<213>artificial sequence
<220>
<223>the VH CDR1 (CDR-H1) of anti-C1s antibody
<400> 36
Gly Phe Asn Ile Lys Asp Asp
1 5
<210> 37
<211> 3
<212> PRT
<213>artificial sequence
<220>
<223>the VH CDR2 (CDR-H2) of anti-C1s antibody
<400> 37
Ala Asp Gly
1
<210> 38
<211> 8
<212> PRT
<213>artificial sequence
<220>
<223>the VH CDR3 (CDR-H3) of anti-C1s antibody
<400> 38
Gly Tyr Gly Arg Glu Val Phe Asp
1 5
<210> 39
<211> 7
<212> PRT
<213>artificial sequence
<220>
<223>the VL CDR1 (CDR-L1) of anti-C1s antibody
<400> 39
Asp Ser Tyr Met Asn Trp Tyr
1 5
<210> 40
<211> 10
<212> PRT
<213>artificial sequence
<220>
<223>the VL CDR2 (CDR-L2) of anti-C1s antibody
<400> 40
Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu
1 5 10
<210> 41
<211> 8
<212> PRT
<213>artificial sequence
<220>
<223>the VL CDR3 (CDR-L3) of anti-C1s antibody
<400> 41
Gln Gln Ser Asn Glu Asp Pro Trp
1 5
<210> 42
<211> 6
<212> PRT
<213>artificial sequence
<220>
<223>the VH CDR1 (CDR-H1) of anti-C1s antibody
<400> 42
Lys Asp Asp Tyr Ile His
1 5
<210> 43
<211> 13
<212> PRT
<213>artificial sequence
<220>
<223>the VH CDR2 (CDR-H2) of anti-C1s antibody
<400> 43
Trp Ile Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys
1 5 10
<210> 44
<211> 11
<212> PRT
<213>artificial sequence
<220>
<223>the VH CDR3 (CDR-H3) of anti-C1s antibody
<400> 44
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp
1 5 10
<210> 45
<211> 107
<212> PRT
<213>artificial sequence
<220>
<223> TNT020 VH3/VK2-Fc-sub4
<400> 45
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
1 5 10 15
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
20 25 30
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
35 40 45
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
50 55 60
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
65 70 75 80
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
85 90 95
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105
<210> 46
<211> 446
<212> PRT
<213>artificial sequence
<220>
<223>VH1 maturation heavy chain
<400> 46
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Leu Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Ala Thr Ile Thr Ala Asp Thr Ser Thr Asn Thr Ala Tyr
65 70 75 80
Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val
260 265 270
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His
420 425 430
Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
435 440 445
<210> 47
<211> 446
<212> PRT
<213>artificial sequence
<220>
<223>VH2 maturation heavy chain
<400> 47
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Ala Thr Ile Thr Ala Asp Thr Ser Thr Asn Thr Ala Tyr
65 70 75 80
Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val
260 265 270
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His
420 425 430
Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
435 440 445
<210> 48
<211> 446
<212> PRT
<213>artificial sequence
<220>
<223>VH4 maturation heavy chain
<400> 48
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val
260 265 270
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His
420 425 430
Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
435 440 445
<210> 49
<211> 446
<212> PRT
<213>artificial sequence
<220>
<223>VH5 maturation heavy chain
<400> 49
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Asp
20 25 30
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Arg Ile Asp Pro Ala Asp Gly His Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Val Lys Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Tyr Gly Arg Glu Val Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val
260 265 270
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His
420 425 430
Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
435 440 445
<210> 50
<211> 218
<212> PRT
<213>artificial sequence
<220>
<223>1 variant maturation light chain of V Kappa
<400> 50
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Thr Gly Gln Pro Pro
35 40 45
Lys Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Glu Glu Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105 110
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
115 120 125
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
145 150 155 160
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
195 200 205
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 51
<211> 218
<212> PRT
<213>artificial sequence
<220>
<223>5 variant maturation light chain of V Kappa
<400> 51
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105 110
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
115 120 125
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
145 150 155 160
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
195 200 205
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 52
<211> 327
<212> PRT
<213>artificial sequence
<220>
<223>4 constant region of human IgG (Fc)
<400> 52
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
1 5 10 15
Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr
65 70 75 80
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro
100 105 110
Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
115 120 125
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
130 135 140
Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
145 150 155 160
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
180 185 190
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu
195 200 205
Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
210 215 220
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys
225 230 235 240
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
245 250 255
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
260 265 270
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
275 280 285
Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
290 295 300
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
305 310 315 320
Leu Ser Leu Ser Leu Gly Lys
325
<210> 53
<211> 327
<212> PRT
<213>artificial sequence
<220>
<223>4 constant region of human IgG (Fc) variant 1
<400> 53
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
1 5 10 15
Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr
65 70 75 80
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro
100 105 110
Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
115 120 125
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
130 135 140
Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
145 150 155 160
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
180 185 190
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu
195 200 205
Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
210 215 220
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys
225 230 235 240
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
245 250 255
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
260 265 270
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
275 280 285
Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
290 295 300
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
305 310 315 320
Leu Ser Leu Ser Leu Gly Lys
325
<210> 54
<211> 218
<212> PRT
<213>artificial sequence
<220>
<223>the anti-C1s VL maturation light chain of parent mouse
<400> 54
Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30
Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Thr Gly Gln Pro Pro
35 40 45
Lys Ile Leu Ile Tyr Asp Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His
65 70 75 80
Pro Val Glu Glu Glu Asp Ala Ala Ile Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
115 120 125
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
145 150 155 160
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
195 200 205
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215

Claims (20)

1. a kind of antibody includes heavy chain and light chain, wherein the heavy chain includes the weight chain variable area (VH) and heavy chain constant region, and this is light Chain includes the area light chain variable (VL)；
Wherein the area VL includes VL complementary determining region (CDR) 1, VL CDR2, and VL CDR3, and wherein the area VH includes VH CDR1, VH CDR2, and VH CDR3；
Wherein the VL CDR1 includes SEQ ID NO:1；
Wherein the VL CDR2 includes SEQ ID NO:2；
Wherein the VL CDR3 includes SEQ ID NO:3；
Wherein the VH CDR1 includes SEQ ID NO:4；
Wherein the VH CDR2 includes SEQ ID NO:5；
Wherein the VH CDR3 includes SEQ ID NO:6；
Wherein the heavy chain constant region includes IgG4 constant region, wherein the amino acid of the heavy chain constant region corresponding to SEQ ID NO:28 Residue 308 is Leu, and the amino acid residue 314 of the heavy chain constant region corresponding to SEQ ID NO:28 is Ser；
And wherein the antibody specificity combines the C1s of activation.
2. the antibody of claim 1, wherein the amino acid residue 108 of the heavy chain constant region corresponding to SEQ ID NO:28 is Pro.
3. the antibody of claims 1 or 2, wherein the amino acid residue 115 of the heavy chain constant region corresponding to SEQ ID NO:28 is Glu。
4. a kind of antibody includes heavy chain and light chain, wherein the heavy chain includes the weight chain variable area (VH) and heavy chain constant region, and this is light Chain includes the area light chain variable (VL)；
Wherein the area VL includes VL complementary determining region (CDR) 1, VL CDR2, and VL CDR3, and wherein the area VH includes VH CDR1, VH CDR2, and VH CDR3；
Wherein the VL CDR1 includes SEQ ID NO:1；
Wherein the VL CDR2 includes SEQ ID NO:2；
Wherein the VL CDR3 includes SEQ ID NO:3；
Wherein the VH CDR1 includes SEQ ID NO:4；
Wherein the VH CDR2 includes SEQ ID NO:5；
Wherein the VH CDR3 includes SEQ ID NO:6；
Wherein the heavy chain constant region includes SEQ ID NO:28；
And wherein the antibody specificity combines the C1s of activation.
5. the antibody of any one of Claims 1-4, wherein the area VL includes and 24 compositions selected from by SEQ ID NO:20,22 The amino acid sequence of group.
6. the antibody of any one of claim 1 to 5, wherein the area VH includes selected from by SEQ ID NO:10,12,14,16, and 18 The amino acid sequence of the group of composition.
7. the antibody of any one of claim 1 to 6, wherein:
(a) it includes SEQ ID NO:20 that the area VH, which includes the area NO:10, Qie Gai VL SEQ ID,；
(b) it includes SEQ ID NO:22 that the area VH, which includes the area NO:10, Qie Gai VL SEQ ID,；
(c) it includes SEQ ID NO:24 that the area VH, which includes the area NO:10, Qie Gai VL SEQ ID,；
(d) it includes SEQ ID NO:20 that the area VH, which includes the area NO:12, Qie Gai VL SEQ ID,；
(e) it includes SEQ ID NO:22 that the area VH, which includes the area NO:12, Qie Gai VL SEQ ID,；
(f) it includes SEQ ID NO:24 that the area VH, which includes the area NO:12, Qie Gai VL SEQ ID,；
(g) it includes SEQ ID NO:20 that the area VH, which includes the area NO:14, Qie Gai VL SEQ ID,；
(h) it includes SEQ ID NO:22 that the area VH, which includes the area NO:14, Qie Gai VL SEQ ID,；
(i) it includes SEQ ID NO:24 that the area VH, which includes the area NO:14, Qie Gai VL SEQ ID,；
(j) it includes SEQ ID NO:20 that the area VH, which includes the area NO:16, Qie Gai VL SEQ ID,；
(j) it includes SEQ ID NO:22 that the area VH, which includes the area NO:16, Qie Gai VL SEQ ID,；
(k) it includes SEQ ID NO:24 that the area VH, which includes the area NO:16, Qie Gai VL SEQ ID,；
(l) it includes SEQ ID NO:20 that the area VH, which includes the area NO:18, Qie Gai VL SEQ ID,；
(m) it includes SEQ ID NO:22 that the area VH, which includes the area NO:18, Qie Gai VL SEQ ID,；Or
(n) it includes SEQ ID NO:24 that the area VH, which includes the area NO:18, Qie Gai VL SEQ ID,.
8. the antibody of any one of claim 1 to 7, it includes SEQ ID that wherein the area VH, which includes the area NO:14, Qie Gai VL SEQ ID, NO:22。
9. the antibody of any one of claim 1 to 8, wherein the light chain further includes constant region of light chain.
10. the antibody of claim 9, wherein the constant region of light chain includes SEQ ID NO:45.
11. the antibody of any one of claims 1 to 10, wherein the heavy chain includes SEQ ID NO:29.
12. the antibody of any one of claim 1 to 11, wherein the light chain includes SEQ ID NO:30.
13. the antibody of any one of claim 1 to 12, is bispecific antibody or multi-specificity antibody.
14. a kind of immunoconjugates of the antibody comprising any one of claim 1 to 13.
15. a kind of nucleotide of the nucleotide set group of the antibody of any one of coding claim 1 to 13.
16. a kind of carrier or carrier set group of the nucleotide of the nucleotide set group comprising claim 15.
17. the place of a kind of nucleotide of nucleotide set group comprising claim 15 or the carrier of claim 16 or carrier set group Chief cell.
18. a kind of antibody comprising any one of claim 1 to 13, the immunoconjugates of claim 14, claim 15 Nucleotide or nucleotide set group, the carrier or carrier set group of claim 16, or the host cell of claim 17, and pharmacy can Receive the pharmaceutical compositions of excipient.
19. a kind of method for inhibiting complement pathway in subject in need, comprising applying pharmacy effective dose to the subject Any one of claim 1 to 13 antibody, the immunoconjugates of claim 14, the nucleotide or nucleotide of claim 15 Set group, the carrier or carrier set group of claim 16, or the host cell of claim 17, or the pharmaceutical composition of claim 18 Object.
20. a kind of method for the disease or illness for treating complement-mediated in subject in need, comprising being applied to the subject With the antibody of any one of a effective amount of claim 1 to 13 of pharmacy, the immunoconjugates of claim 14, the core of claim 15 Thuja acid or nucleotide set group, the carrier or carrier set group of claim 16, or the host cell of claim 17, or claim 18 pharmaceutical compositions.
CN201780074920.XA 2016-10-12 2017-10-12 anti-C1 s antibodies and methods of use thereof Active CN110300520B (en)

Priority Applications (1)

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